To examine whether socioeconomic position (SEP) is associated with progression in the health-seeking process for hearing loss.Logistic regression of data from a cross-sectional survey representative of noninstitutionalized, 50 years and older population of England . Using self-reported hearing difficulty as starting point, we examined the association between SEP and health-seeking behaviors in 6 stages leading to hearing aid acquisition and use.p < .001). There was marginal negative association between higher SEP and receiving hearing aid recommendation . SEP was not associated with any other stage of health-seeking behavior.Higher SEP was associated with lower odds of self-reported hearing difficulty, adjusted odds ratio [OR] = 0.87 (95% confidence interval [CI] 0.83–0.91, Among the noninstitutionalized older population of England, SEP-related inequalities exist in the prevalence of self-reported hearing loss. However, SEP is not strongly associated with progression in the remaining stages of health-seeking process during and after an individual’s contact with the health system. Health-A formal comparison of causal mechanisms linking socioeconomic position (SEP) and health outcomes among the older population in England showed that financial resources and behaviors, rather than psychosocial factors, were significantly associated with gradients in health status . TherefoCurrent research about socioeconomic determinants of health seeking for hearing loss is limited mainly to examination of hearing aid ownership. A recent systematic literature review did not identify any studies examining the association between SEP and likelihood of reporting hearing difficulty to a health professional . Among oUntreated hearing difficulty can negatively impact on mental and physical health, yet hearing impairment among older individuals is underdiagnosed and undertreated in the United Kingdom . We wereThis study used data from the second wave of the ELSA due to availability of self-reported information on the existence of hearing difficulty and consecutive multiple stages of health-seeking behavior. ELSA core members are a representative sample of the noninstitutionalized population who were 50 years or older on March 1, 2002, and living in England. The ELSA cohort originated from the Heath Survey for England (HSE or ELSA Wave 0), which provided enrollment of more than 12,000 core members and baseline data about them between years 1998 and 2001 . The samRespondents were asked to rate their hearing on a scale from excellent, very good, good, fair to poor. In addition, they were asked whether they had difficulties following a conversation in a setting with background noise. If respondents rated their hearing as fair or poor, or acknowledged having difficulties following a conversation, they were asked whether they had mentioned their hearing problems to a health professional (physician or nurse). An affirmative response led to being asked whether the respondents were referred to an ear specialist. All individuals who mentioned their hearing difficulty to a health professional were asked whether they received a recommendation for a hearing aid and if so, whether they obtained one. The section concluded by asking whether the respondent uses their hearing aid. These six questions were considered discrete stages in the health-seeking process, and binary responses to them were analyzed separately.All stages of health-seeking behavior, starting with perception of hearing difficulty, were self-reported and not confirmed by other methods. The reliability of the self-reported measure of hearing loss among older people compared to audiometry ranges between populations, depends on severity of hearing impairment, and may vary by demographic characteristics. In Taiwan, 21.4% of older people with moderate to profound hearing impairment (>40 dB HL as measured by audiometry) self-reported hearing difficulty . Among oEducation, occupation, income, and wealth were selected as the four indicators capturing most of the variation in socioeconomic situation  and usedGender and age  were used in the analysis, due to their association with prevalence and extent of hearing impairment . In addip value equalled <.05 in any of the six models, the modification effect was examined and reported.Data were analyzed using Stata/SE v.12 . At each of the six stages of health-seeking behavior, a single model was estimated incorporating the continuous latent measure of SEP and adjusted for gender, age group, current marital status, retirement status, and ownership of private health insurance. Based on previous findings of the importance of gender and cohort effects in the pathways linking socioeconomic status with health outcomes this population , gender,Prevalence of self-reported hearing difficulty was estimated taking the sampling weights and design into account using the Stata “svy” command. Bootstrapping with 1,000 replications was used in each model, and the results of final adjusted models were confirmed using logistic regression including sample weights. The largest percentage of records with missing data in any of the variables was recorded at the first stage (6.3%), and due to low proportion of missingness (<5.7%) in the consecutive stages, records with missing data were eliminated from analysis.Men comprised 45% of the initial sample of 8,780 . Nearly In the adjusted model, self-reported hearing difficulty was strongly associated both with gender and with age group , model 1p = .098). Additionally, the results revealed that individuals who were referred to an ear specialist had 19 times higher odds of receiving a hearing aid recommendation  compared with those who were not referred.In model 3, we did not observe an association between SEP and the odds of being referred to an ear specialist upon mentioning hearing difficulty to a health professional. There was strong evidence that people in the oldest age group had more than double the odds of being referred to an ear specialist. Among individuals who mentioned their hearing difficulty to a health professional , there was some evidence that an increase in SEP score was associated with a decrease in the odds of receiving a recommendation for a hearing aid (model 4). There was strong evidence that the odds of hearing aid being recommended increased with increasing age group. No interaction between age group and gender or age group and SEP was identified. Model 4 was further adjusted for referral to an ear specialist as a potential indicator capturing some of the variation related to the severity of hearing loss. This analysis showed that SEP was no longer associated with the odds of receiving a hearing aid recommendation , whereas this effect was not seen in the older two age groups  women had 2.6 times higher odds of acquiring a hearing aid compared with men , and willingness to report its existence to the survey enumerator. Different types of bias may have occurred in self-reporting hearing loss, receiving a hearing aid recommendation and compliance with this recommendation. Because the objective severity of hearing impairment was not available, we could not adjust for it in our models. Although the sample analyzed is derived from a longitudinal cohort, detailed information about health-seeking behavior for hearing loss was available only in Wave 2. The cross-sectional nature of our analysis did not allow for information about the date and length of the health-seeking process  to be considered.However, we believe it is unlikely that the existence of hearing impairment could fully explain the existence of the gradient in self-reported hearing handicap, in light of research showing that there was no evidence that social mobility was caused by health status . The useAmong the older population in England, nearly half of self-perceived hearing difficulty is unreported to health professionals, and therefore remains untreated. SEP was strongly associated with odds of self-reported hearing difficulty. In light of a high prevalence of hearing difficulty and absence of a hearing loss screening program among older individuals in the United Kingdom, the strong associations between self-reported hearing difficulty and increasing age, male gender, and lower SEP may aid health practitioners in raising the issue with particular patients during other health care interactions. In regard to the objective of this study, we found that individuals of different SEP scores appeared to receive equitable levels of care upon approaching a health care professional with self-perceived hearing loss, and no SEP gradient was identified in ownership and utilization of hearing aids.Women with a self-perceived hearing handicap were less likely to discuss its existence with a health professional, but more likely to obtain a hearing aid upon recommendation. Further research is needed to evaluate how women may be encouraged to report this sensory impairment and how uptake of hearing aids could be improved among men, especially in the youngest age group. Whereas the current system does not appear to disadvantage individuals from lower SEP backgrounds, any changes to screening or treatment availability and eligibility will need to assess the potential for creating such gradients.In addition to the above recommendations targeted at clinical practice, we offer two potential avenues of future research to help elucidate the pathways linking SEP and health outcomes via health-seeking behaviors. Firstly, comparative country studies of accessibility and affordability of hearing loss care in various health system environments may assist in designing interventions to lower barriers to receiving appropriate treatment. Secondly, a more general exploration of the influence of health-seeking behavior in the causal framework between socioeconomic behavior and health outcomes is needed. A high proportion of hearing impairment is undiagnosed and untreated in the older population in England, and efforts need to be made to screen and encourage the use of hearing aids in order to prevent avoidable morbidity in this population. To guide effective policy design, future analysis needs to simultaneously explore the relative effects of all causal pathways  linking SEP with health outcomes.Economic and Social Research Council (ESRC) 1+3 Studentship grant award (ES/I903224/1) and a Medical Research Council (MRC) Population Health Science fellowship (G0802442).This work was supported by the


However, further investigation is required to unequivocally establish the absence of correlation between the platelet parameters and occurrence of SSNHL.Sudden sensorineural hearing loss (SSNHL) is a common otological disorder characterized by a hearing loss greater than 30 dB over three consecutive frequencies, in less than 72 hours. It has been established that platelet parameters, such as mean platelet volume, are associated with ischemic heart events, whose clinical manifestations are similar to those of SSNHL. Hence, we aimed to determine if the platelet count, mean platelet volume and platelet distribution width are related to the occurrence and severity of sudden sensorineural hearing loss. A case-control prospective study was conducted in a teaching hospital in Iran. One hundred-eight patients with SSNHL and an equal number of healthy, age- and sex-matched controls were enrolled in the study. Peripheral venous blood samples were collected from the subjects, and the platelet count, mean platelet volume and platelet distribution width were measured with an automated blood cell counter. Analysis of the audiometry and hematological test results using SPSS Hearing loss is a common, escalating disorder worldwide, which has multiple negative consequences on the afflicted. Studies have shown that about 50% of people are affected by hearing loss, which increases with age. The occurrence of hearing loss has seen an increase in the past decades, especially in developing countries, with its incidence doubling between 1965 and 1994 .Sudden sensorineural hearing loss (SSNHL) is a subset of sudden hearing loss that has a sensory-neural origin . SSNHL, Several reports have detailed the prevalence of SSNHL in various countries ,8–12. ReSudden sensorineural hearing loss occurs at all ages, with childhood incidences being rare. The lowest incidence in adults has been reported to be among the age group of 20–30, and the highest among 50–60 ,8. The pSudden sensorineural hearing loss presents symptoms such as tinnitus, dizziness with 10% of these patients experiencing dizziness with nausea and vomiting, fullness of the ear, a sense of ear congestion, vestibular disorders, feeling of pressure in the ears, headaches, and symptoms of a viral infection of the upper respiratory tract. Patients also might complain of anxiousness, stress and depression ,11,18,19Many aspects of SSNHL are still unknown, and its etiology, risk factors, prognostic factors and treatment protocols remain controversial ,20. ProbMean platelet volume (MPV) is a potential indicator of the production rate, size, activity, and function of platelets. Larger platelets are more active in terms of both metabolic and enzymatic activity, and more likely to aggregate than the smaller platelets ,14,31. SThe aim of this study was to investigate the relationship of platelet parameters  with the occurrence and severity of sudden sensorineural hearing loss. Therefore in this study we chose to analyze the following variables:Sudden sensorineural hearing loss (SSNHL): We chose this variable for our study because although SSNHL is a common otological disorder with an increasing rate of incidence worldwide but to date its many aspects are still unknown and need to be studied more.Platelets parameters including PC, MPV and PDW: We chose to analyze the role of platelets parameters on SSNHL because some authors have claimed that the occurrence and severity of SSNHL could be correlated with platelets parameters due to its similar clinical manifestation to that of vascular diseases. However, so far this likely correlation has been investigated in few studies to our knowledge. So, we think that it is a research area where we need more information.A prospective case-control study was conducted in the Shahid Sadoughi hospital, Yazd, Iran, between 2013 and 2015. One hundred-eight patients with SSNHL and the same number of healthy age- and sex-matched subjects were enrolled as subjects for the study. The study population consisted of patients referred to the ENT department of hospital. An informed written consent in the native language (Persian) was obtained from all the participants in the study, and the researchers were committed to maintaining confidentiality of participant information. Approval for this study was obtained from the Medicine School of Shahid Sadoughi University of Medical Sciences, Yazd, Iran, the ENT department of Shahid Sadoughi hospital, Yazd, Iran and the ethical committee of Shahid Sadoughi University of Medical Sciences, Yazd, Iran.Patients with inflammation, infection, renal failure, chronic liver disease, pulmonary thromboembolism, chronic obstructive pulmonary disease (COPD), systemic hypertension, cardiovascular disease, diabetes mellitus, metabolic syndrome, hyperlipidemia, statin usage, obstructive sleep apnea, rheumatological diseases, inflammatory bowel disease, hematological or endocrine diseases, history of smoking, otological disease or surgery or trauma to the ear or head, were excluded from the study.An audiometry evaluation was performed for the chosen test subjects. All evaluations were performed by the same hearing therapist, and with the same audiometer. Audiometry evaluations included pure-tone audiometry (PTA), speech audiometry (including SDS and SRT), and Immittance criteria (including tympanometry and stapedius reflex).Pure-tone audiometry is the standard and most common type of hearing test which measures the air and bone conduction thresholds for each ear in a set of standard frequencies from 250hz to 8000hz. The test is conducted in a sound booth using a pair of headphones connected to an external audiometer. The result of the test is an audiogram diagram which plots a person's hearing sensitivity at the tested frequencies. In this study, first, each patient was instructed about the procedure. Then the earphone was placed on the ear and checked for secure and proper fit. Then the pure tones at different frequencies were presented to the patient from the low to high intensity and the lowest audible intensity was defined as the patient’s threshold for that particular frequency which is marked as such on the audiogram. This procedure was repeated for all test frequencies  in one ear and then again in the other ear. This procedure established an air conduction pure tone threshold curve for each ear. Then the bone conduction audiometry was done. For this, the probe was placed on the mastoid, the same procedure as the air conduction audiometry was done and the bone conduction pure tone threshold curve was established for each ear.SRT (speech reception threshold) which also called speech recognition threshold is the minimum intensity in decibels at which a patient can understand 50% of spoken words and can repeat them correctly. In this test, a number of spondaic words are presented to the patient at different intensities. Spondaic words, or spondees, are words containing two syllables that are equally accented or emphasized when they are spoken to the patient. In this study, we presented 4 spondaic words to each patient at different intensities and the lowest intensity in which the patient repeated 50% of presented spondees correctly was defined as SRT. The SRT was recorded in db for each ear, separately.Speech discrimination testing, also known as “word understanding” or “word recognition” is another important audiological test. The aim of this test is to assess how well an individual can understand words which are fully audible based on the level of hearing (as measured via SRT). In this study, 25 unfamiliar one-syllable words were presented to each patient at 40 db above SRT. The percentage of words which patient understood and repeated correctly was recorded as Speech Discrimination Score (SDS) for each ear separately.Tympanometry is an examination used to test the condition of the middle ear and mobility of the eardrum (tympanic membrane) and the conduction bones by creating variations of air pressure in the ear canal. It is an objective test of middle-ear function. It is not a hearing test, but rather a measure of energy transmission through the middle ear. Tympanometry is a valuable component of the audiometric evaluation. In evaluating hearing loss, Tympanometry permits a distinction between sensorineural and conductive hearing loss, when evaluation is not apparent via Weber and Rinne testing. In this test, a tone of 226 Hz is generated by the Tympanometer into the ear canal, where the sound strikes the tympanic membrane, causing vibration of the middle ear, which in turn results in the conscious perception of hearing. Some of this sound is reflected back and picked up by the instrument. Most middle ear problems result in stiffening of the middle ear, which causes more of the sound to be reflected back. Admittance is how energy is transmitted through the middle ear. The instrument measures the reflected sound and expresses it as admittance or compliance, plotting the results on a chart known as a Tympanogram. Normally, the air pressure in the ear canal is the same as ambient pressure. Also, under normal conditions, the air pressure in the middle ear is approximately the same as ambient pressure since the eustachian tube opens periodically to ventilate the middle ear and to equalize pressure. In a healthy individual, the maximum sound is transmitted through the middle ear when the ambient air pressure in the ear canal is equal to the pressure in the middle ear. In our study, after an examination of the ear with an otoscope to ensure that the path to the eardrum is clear and that there is no perforation, the test was performed by inserting the Tympanometry probe in the ear canal of patient. The instrument changed the pressure in the ear, generated a pure tone, measured the eardrum responses to the sound at different pressures and produced a Tympanogram.The acoustic reflex  reflex, attenuation reflex, or auditory reflex) is an involuntary muscle contraction that occurs in the middle ear in response to high-intensity sound stimuli or when the person starts to vocalize. This reflex is a normal defensive mechanism for inner ear against the loud sounds. The acoustic reflex threshold (ART) is the sound pressure level from which a sound stimulus with a given frequency will trigger the acoustic reflex which can be measured in each ear with Tympanometry. In our study, the stapedius reflex was tested with Tympanometry for each ear of each patient and the acoustic reflex threshold was recorded.In this study, after the performing of audiometry evaluation, the subjects who had developed SSNHL within 72 hours were entered into the study as the patient group. All the patients had a minimum 30 dB hearing loss at three consecutive frequencies. For analyzing the severity of hearing loss according to platelet parameters, the patient group was divided into 4 subgroups based on the severity of hearing loss as mild (<40 db), moderate (40–55 db), sever (55–90 db) and profound (>90 db) hearing loss. The control group participants were selected from the non-otologic patients of the ward. The exclusion criteria for the control subjects were similar to those of patient group. Therefore, patients with inflammation, infection, renal failure, chronic liver disease, pulmonary thromboembolism, chronic obstructive pulmonary disease (COPD), systemic hypertension, cardiovascular disease, diabetes mellitus, metabolic syndrome, hyperlipidemia, statin usage, obstructive sleep apnea, rheumatological diseases, inflammatory bowel disease, hematological or endocrine diseases, history of smoking or surgery or trauma to the ear or head were excluded from the control group.22  software. The data were summarized using descriptive statistics. Box and scatter plots were used to present descriptive statistics. An independent sample T-test, ANOVA, Pearson correlation coefficient and regression analysis were used to analyze the data. A two-tail value of p<0.05 was considered statistically significant.Venous blood samples were collected from all the participants in tubes containing Ethylenediaminetetraacetic acid (EDTA) at 7:30 am, following an overnight fast. Mean platelet volume (MPV), platelet distribution width (PDW) and platelet count (PC) were measured for each of the blood samples. All measurements were performed with an automated blood cell counter, within 15–30 minutes after sampling, in order to avoid platelet swelling. All measurements were carried out in duplicates, and the mean values were analyzed. Data analysis was conducted using the SPSSThe male:female ratios of the two groups were 61:47 . The difThe platelet parameters that were tested showed no significant variation between the patient and control groups. The clinical laboratory results of patients and control groups including the values of PC, MPV and PDW are presented in As shown in As presented in Also, the analysis of platelet parameters according to severity of hearing loss is presented in For more clarity, we draw the scatterplots of platelets parameters versus severity of hearing loss. Figs As shown in As presented in the above table, PDW showed significant correlation with hearing loss which is seen in scatterplot 3, too.Sudden sensorineural hearing loss is a common otological disorder with an increasing rate of incidence worldwide, based on current reports. SSNHL requires immediate attention due to its varied etiologic factors and prognostic features ,34. AlthDue to the diverse etiology of SSNHL, its treatment has not been satisfactory . In addiPrevious studies have shown that platelets parameters such as mean platelet volume are associated with ischemic heart events , and in Few studies have examined the relationship of these variables with sudden sensorineural hearing loss. Seo et al. 2014), in a study to identify diagnostic and prognostic biomarkers in patients with SSNHL, compared the biomarker profile of patient and healthy control groups, and found that they differed significantly in their platelet count . However, in a stS1 Table(DOC)Click here for additional data file.S2 Table(DOC)Click here for additional data file.


The International Classification of Functioning, Disability and Health (ICF) proposes that there are three primary healthcare outcomes: body functions/structure, activity and participation . The ICFSemi-structured interviews were conducted with nine hearing healthcare professionals and 25 adults with mild-to-moderate hearing loss. Maximum variation sampling facilitated the recruitment of participants with a variety of characteristics and experiences . The datThe results show that hearing loss can lead to participation difficulties in various social domains, including work, friendship and family. Most of the interviewees with hearing loss successfully participated in some social domains, but struggled in other social domains. This was particularly problematic when they struggled in the social domains they most valued, such as family life. Some reported that, although they had many social interactions, they felt isolated during those interactions. Hearing loss can cause participation difficulties by leading to fatigue, embarrassment and diminished self-confidence in social interactions.This research demonstrates that it is important to measure individuals’ active participation in the social domains that they most value. This contrasts with many previous participation measures that assess how often people participate or the number of social domains in which people participate . The nex


Cathepsin L [EC 3.4.22.15] is secreted via lysosomal exocytosis by several types of cancer cells, including prostate and breast cancer cells. We previously reported that human cultured fibrosarcoma (HT 1080) cells secrete cathepsin L into the medium; this secreted cathepsin is 10-times more active than intracellular cathepsin. This increased activity was attributed to the presence of a 32-kDa cathepsin L in the medium. The aim of this study was to examine how this active 32-kDa cathepsin L is secreted into the medium. To this end, we compared the secreted active 32-kDa cathepsin L with lysosomal cathepsin L by using a novel gelatin zymography technique that employs leupeptin. We also examined the glycosylation and phosphorylation status of the proteins by using the enzymes endoglycosidase H [EC 3.2.1.96] and alkaline phosphatase [EC 3.1.3.1]. Strong active bands corresponding to the 32-kDa and 34-kDa cathepsin L forms were detected in the medium and lysosomes, respectively. The cell extract exhibited strong active bands for both forms. Moreover, both forms were adsorbed onto a concanavalin A-agarose column. The core protein domain of both forms had the same molecular mass of 30 kDa. The 32-kDa cathepsin L was phosphorylated, while the 34-kDa lysosomal form was dephosphorylated, perhaps because of the lysosomal marker enzyme, acid phosphatase. These results suggest that the active 32-kDa form does not enter the lysosomes. In conclusion, our results indicate that the active 32-kDa cathepsin L is secreted directly from the HT 1080 cells and not via lysosomal exocytosis. This lysosomal docking and fusion are regulated by a transcription factor EB (TFEB) for lysosome biogenesis and autophagy  and L [EC 3.4.22.15] are cysteine proteinases belonging to the papain family that cleave the extracellular matrix when secreted outside the cells. Several independent studies have reported the importance of cathepsins in cancer invasion and metastasis n cancer ; increasn cancer ,12. Cancn cancer ,12; howeN-linked oligosaccharides; the mannose moiety of cathepsin L is phosphorylated by UDP-N-acetylglucosamine:lysosomal enzyme N-acetylglucosaminylphosphotransferase [EC 2.7.8.17] in the cis Golgi ) and alkaline phosphatase [EC 3.1.3.1] (ALP). Our results provide insights into the molecular mechanism underlying the membrane trafficking of the active 32-kDa cathepsin L form secreted by HT 1080 cells.In the present study, we sought to investigate the mechanism underlying the secretion of the active 32-kDa cathepsin L form into the medium by HT 1080 cells. The secretion mechanism was examined by comparing the secreted and lysosomal cathepsin L forms using the improved gelatin zymography method and by employing two enzymes, endoglycosidase H [EC 3.2.1.96] . ALP (P 4245) was purchased from Sigma-Aldrich Co. . Prestained SDS-PAGE standard (broad range) was purchased from Bio-Rad Laboratories . Lectin-agarose minicolumn Kit I, which was packed with concanavalin A , Lens culinaris agglutinin , Ricinus communis agglutinin 120 , and wheat germ agglutinin , was purchased from Seikagaku Biobusiness Co. . CA-074 and CLIK-148 were synthesized by Katunuma’s group [Leupeptin and Z-Phe-Arg-MCA were purchased from the Peptide Institute . Dulbecco’s modified Eagle’s medium (DMEM) and fetal calf serum (FCS) were purchased from Invitrogen Corp. . The human fibrosarcoma cell line HT 1080 was purchased from Dainippon Pharmaceutical Co., Ltd. . End H from 5 cells) were seeded in Falcon™ culture dishes  and cultured in DMEM in the presence of 200 U/mL penicillin, 0.4 mg/mL streptomycin, 0.2 mg/mL kanamycin, 2 μL/mL Fungizone, and 10% FCS. The cells were then passaged at confluence (up to the fifth generation) at 37°C under 5% CO2 in air. After reaching confluence, the cells were washed with serum-free DMEM, and then cultured in the same medium for 24 h. The culture medium was removed, and the cells were washed twice with 1 mL of phosphate-buffered saline (PBS) and collected by centrifugation at 700 ×g for 10 min. Enzymes were extracted from the cells by homogenization in 20 mM Tris-HCl buffer, pH 6.8. After centrifugation at 11,400 ×g for 10 min at 4°C, the resultant supernatant (cell fraction) and culture medium were stored at −80°C.HT 1080 cells  and/or cathepsin L (5 μM CLIK-148) in 50 mM sodium acetate buffer (pH 5.5) for 10 min at 37°C. The amount of 7-amino-4-methylcoumarin liberated from the substrate was estimated fluorometrically in a Shimadzu RF-540 fluorescence spectrometer  at excitation and emission wavelengths of 370 nm and 460 nm, respectively. The net activity was calculated by subtracting the activity in the presence of inhibitors of cathepsin L and cathepsin B from that in the presence of cathepsin B, as described by Inubushi et al. . One uniThe gelatin-degrading activities were examined on 11.5% polyacrylamide gels containing 0.8 mg/mL gelatin, using a slab gel measuring 85 × 80 × 1 mm, as described previously ,23. The Carbohydrate species attached to the secreted and lysosomal cathepsin L forms were examined using a Lectin-agarose minicolumn Kit I packed with Con A, LCA, RCA120, and WGA, according to the manufacturer’s instructions. Briefly, 400 μL of sample, which was prepared by diluting both cathepsin forms with PBS containing 0.05 mM leupeptin (column buffer), was loaded onto each lectin-packed minicolumn pre-equilibrated with column buffer. The column was then washed with 2 mL of column buffer (this eluate was pooled as the non-adsorption fraction), and the adsorbed materials were eluted from the column with 10 mM methyl-α-D-glucoside for the Con-A agarose column (this eluate was pooled as the adsorption fraction). Both forms were concentrated by using Centricon-10 , and gelatin zymography was performed to examine which fraction contained the 32- and/or 34-kDa cathepsin L.The carbohydrate moiety was removed from the secreted 32-kDa and lysosomal cathepsin L forms by End H treatment as follows: End H  was solubilized with 100 μL of 10 mM Tris-HCl buffer (pH 7.2) containing 0.1% bovine serum albumin . An aliquot of the cell fraction, lysosomal fraction, or culture medium was mixed with or without 10 mU of End H in 0.1 M sodium acetate buffer (pH 5.0) containing 0.1 mM leupeptin, and then incubated for 30 min at 37°C. Gelatin zymography was performed for each incubation mixture.2, 10 μM ZnCl2, and 0.1 mM spermidine, and incubated for 30 min at 37°C. Gelatin zymography was performed for each incubation mixture.An aliquot of the cell fraction, lysosomal fraction, or culture medium was mixed with or without ALP in 0.5 mM Tris-HCl buffer (pH 9.3 at 25°C) containing 0.1 mM leupeptin, 0.1 mM MgClA portion of the lysosomal fraction or culture medium was incubated in 0.1 M sodium acetate buffer (pH 4.8) for 0, 2, or 4 h at 37°C in the presence or absence of leupeptin. After incubation, aliquots of the incubation mixtures were subjected to gelatin zymography to examine whether or not the secreted and lysosomal cathepsin L forms were processed further.HT 1080 cell fraction  and culture medium  were subjected to gelatin zymography in the presence or absence of leupeptin. As shown in Gelatinolytic activity (with an optimum pH of 4.8) in all forms required thiol reagents and was inhibited by cysteine proteinase inhibitors such as E-64, leupeptin, or N-acetyl-Leu-Leu-methional, but not by ethylenediaminetetraacetic acid, an inhibitor of metalloproteinases, phenylmethylsulphonylfluoride, an inhibitor of serine proteinases, or pepstatin A, an inhibitor of aspartic proteinases . MoreoveN-linked high-mannose-type oligosaccharides.Both the 32- and 34-kDa forms were detected in the adsorption fraction of the Con A-agarose column . In the Lens culinaris agglutinin), RCA120 (Ricinus communis agglutinin 120), and WGA (wheat germ agglutinin), as described in the Materials and Methods. Our improved gelatin zymography method was performed to examine whether the 32- and/or 34-kDa forms were bound to the lectins.Oligosaccharide species bound to the 32- and/or 34-kDa cathepsins L were examined using a Lectin-agarose minicolumn Kit I packed with Con A , LCA  are transported to lysosomes via endosomes . The seqnal side ,37. The nal side . Subsequnal side . This exKuliawat et al.  reportedInhibition of invasion and metastasis is the method-of-choice for the treatment of cancers. Many cancer cells secrete active cathepsins into the extracellular environment. The secreted cathepsins then degrade the extracellular matrix, and enable invasion and metastasis in the acidic microenvironment. Therefore, studies investigating the mechanisms underlying the secretion of active cathepsins by cancer cells may lead to the development of novel cancer therapies.In conclusion, our results suggest that the active 32-kDa cathepsin L form detected in the culture medium is secreted directly from the HT 1080 cells and not via lysosomal exocytosis or extracellular processing of procathepsin L.


The visual system is beautifully crafted to transmit information of the external world to visual processing and cognitive centers in the brain. For visual information to be relayed to the brain, a series of axon pathfinding events must take place to ensure that the axons of retinal ganglion cells, the only neuronal cell type in the retina that sends axons out of the retina, find their way out of the eye to connect with targets in the brain. In the past few decades, the power of molecular and genetic tools, including the generation of genetically manipulated mouse lines, have multiplied our knowledge about the molecular mechanisms involved in the sculpting of the visual system. Here, we review major advances in our understanding of the mechanisms controlling the differentiation of RGCs, guidance of their axons from the retina to the primary visual centers, and the refinement processes essential for the establishment of topographic maps and eye-specific axon segregation. Human disorders, such as albinism and achiasmia, that impair RGC axon growth and guidance and, thus, the establishment of a fully functioning visual system will also be discussed. The eyes together with their connecting pathways to the brain form the visual system. In the eye, the cornea bends light rays and is primarily responsible for focusing the image on the retina. The lens behind the cornea inverts the image top to bottom and right to left. The retina, the receptive surface inside the back of the eye, is the structure that translates light into nerve signals, and enables us to see under conditions that range from dark to sunlight, discriminate colors, and provide a high degree of visual precision. The retina consists of three layers of nerve cell bodies separated by two layers containing synapses made by the axons and dendrites of these cells. The back of the retina comprises the photoreceptors, the rods, and cones. The medial retinal layer contains three types of nerve cells, bipolar, horizontal, and amacrine cells. Bipolar cells receive input from the photoreceptors, and many of them feed directly into the retinal ganglion cells (RGCs). Horizontal cells connect receptors and bipolar cells by relatively long connections that run parallel to the retinal layers. Amacrine cells link bipolar cells and RGCs, the cells located in the inner retina. RGC axons pass across the surface of the retina and are collected in a bundle at the optic disk to leave the eye and form the optic nerve. There are approximately 20 RGC types that can be classified by morphological, molecular, and functional criteria. Each RGC type participates in distinct retinal circuits and projects to a specific set of targets in the brain , includiThe optic axons from both eyes meet at the optic chiasm, which is located at the base of the hypothalamus. There, RGC axons from the nasal retina cross over to the opposite side of the brain  while axons from the temporal retina turn to project to the same hemisphere as their side of origin . Through evolution, the extent of binocular vision correlates with the frontalization of the eyes and, consequently, the proportion of ipsilateral axons . In primA vast number of axons of the optic tract terminate in the LGN. The right LGN receives information from the left visual field , while the left LGN receives information from the right visual field  is essential for commitment toward the RGC fate. Atoh7 is expressed in retinal progenitors coincident with the onset of RGC differentiation. In mice lacking Atoh7, few RGCs are generated, with progenitor cells instead adopting an amacrine or cone photoreceptor fate  surface, forming spherical cells with no obvious polarity. These apolar cells extend a basal process through the thickness of the retina toward the vitreal surface, generating cells with a bipolar morphology attached to the inner and outer limiting membranes of the retina. The RGC nucleus then migrates through the basal process toward the inner surface of the retina. Whilst this migration is occurring, an axon extends, usually from the basal process, into the optic fiber layer. Finally, the bipolarly shaped cell breaks the attachment of its apical process with the ventricular surface, resulting in a morphologically distinguishable RGC located within the RGC layer at the inner surface of the retina. Formation of dendrites occurs after axon initiation but before target innervation .Figure slit1b or robo3, retraction of the apical process and nuclear migration are delayed  surface of the retina  surface of the retina—the optic fiber layer. Growth of retinal axons is restricted to the optic fiber layer through a combination of the growth-promoting properties of the glial endfeet present in this region, and the inhibitory properties of the outer retinal layers . Purifieg retina , or theie retina , 2009. BSlit1/2 or Robo2 mouse mutants  that help drive growth centrally. Blocking the function of these molecules in chick, fish, or rodent retinas impairs the tight bundling together of RGC axons and the disk-directed growth of some axons  and integrin-linked kinase (ILK) are required for the inhibitory signaling of high concentrations of Shh in RGC axons (Hip1 (hedgehog-interacting protein) is also expressed by all RGCs, but it is unclear whether it plays a role in retinal axon directionality.Acting in concert with the inhibitory forces me axons . In zebrtic disk . Anotherntration . Both ovntration . Similarntration . Howeverntration . HoweverGC axons , but thiGC axons . Hip1 (hBrn3b or expressing multiple copies of human PAX6 (+/+Pax77 mice), RGC axon fasciculation is impaired, with many axons following abnormal trajectories within the retina, and, in the case of Brn3b mutants, failing to reach the optic disk  on the RGC axons . Netrin- the eye . At the  the eye . Reverse the eye .Vax1, RGC axons stall as they enter the brain and fail to form the chiasm  rescues RGC axon pathfinding errors in zebrafish mutants with partial loss of Robo2 function  midline, except for a small gap where the optic chiasm develops. Thus, RGC axons normally extend across the midline through a Shh-free region, with high levels of Shh bordering the chiasm both anteriorly and posteriorly . This gaaterally . Consist signals . Blockinic nerve . Thus, sic nerve . WhetherIn all species, the majority of RGC axons project contralaterally at the optic chiasm. However, for many years, the mechanisms that control crossing at the chiasm remained elusive. Chemoattractants, such as netrin-1  and Shh in vitro experiments demonstrated that, when acting alone, NrCAM is required exclusively within the retina for promotion of RGC axon growth on chiasm cells  binding isoform of the classical vascular growth factor VEGF-A . Strong sema3d in zebrafish results in axons stalling in the proximal optic tract and aberrant ipsilateral projections  reduces levels of Nrp1a and Nrp1b mRNA in RGCs. This results in RGC axons guidance errors that phenocopy knockdown of Nrp1a, or its known chiasm ligands in zebrafish, sema3d, and sema3e  function in a semi-intact preparation of the developing visual system results in a reduction in the size of the ipsilateral projection. No effect was observed on the contralateral projection after Nogo downregulation nhibited . NogoR ibelladona/lhx2 mutants. In these mutants, RGCs axons fail to cross the midline due to defects in the organization of the midline glia, with the expression of Sema3d being absent from the diencephalon. In addition, expression of Slit2 is expanded across the midline . EphrinAs on RGC axons also interact with another BDNF receptor, p75  or partially  reversed in the laminae of the thalamus . These atyrosinase gene or other components of the melanin biosynthetic pathway (belladona zebrafish mutants (Lhx2 mutants) exhibit a phenotype similar to achiasmatic individuals with misrouting of retinal axons to the ipsilateral side of the brain accompanied by congenital nystagmus (At the molecular level, albinism is caused by alterations in the expression levels of the  pathway . Some st pathway . In fact pathway , and it  pathway  that is  pathway . Howeverystagmus .At the LGN, misrouted axons from the VT retina of albino mice project within the contralateral LGN forming a patch separated from the contralateral termination area . This reThe abnormal visual input perceived by achiasmatic or albino individuals does not induce a topographic reorganization in the thalamocortical projection or the occipital callosal connections. Instead, albino and achiasmatic individuals present atypical organization of the visual cortex consisting of overlapping visual hemifield maps with bilateral population receptive fields . Thus, rIt is clear from the studies discussed in this review that the wiring together of the developing visual system involves a complex interplay of genetic, molecular and activity based mechanisms. Visual function is critically dependent on the correct specification and generation of RGCs, and appropriate guidance of their axons to visual target regions in the brain. Guidance of RGC axons is not a simple process but requires integrated interactions between multiple coexpressed signals and modulatory factors, as well as regulation of intrinsic changes in growth cone responses and guidance cue expression. Once axons reach their targets, their task is far from complete, and molecular mechanisms act in concert with spontaneous activity to induce rearrangement and refinement of axon termini. Disruption of any of these processes can result in severe visual impairment. Although our understanding of the mechanisms controlling visual system wiring has increased substantially over the past 10 to 20 years, much still remains to be established and will form a significant remaining challenge in the years to come.


C-approximating posets, the concept of countably QC-approximating posets is introduced. With the countably QC-approximating property, some characterizations of generalized completely distributive lattices and generalized countably approximating posets are given. The main results are as follows: (1) a complete lattice is generalized completely distributive if and only if it is countably QC-approximating and weakly generalized countably approximating; (2) a poset L having countably directed joins is generalized countably approximating if and only if the lattice σc(L)op of all σ-Scott-closed subsets of L is weakly generalized countably approximating.As a generalization of countably  L is continuous if and only if the lattice σ(L)op of all Scott-closed subsets of L is completely distributive. Gierz et al. in [C-continuous lattices. And they showed that for any poset L, σ(L)op is a C-continuous lattice and that L is continuous if and only if σ(L)op is continuous.The notion of continuous lattices as a model for the semantics of programming languages was introduced by Scott in . Later, t al. in  introduct al. in  introduct al. in  proved tt al. in  introducσ-Scott topology. Yang and Liu in [C-approximating posets and showed that the lattice of all σ-Scott-closed subsets of a poset is a countably C-approximating lattice and that a complete lattice is completely distributive if and only if it is countably approximating and countably C-approximating.On the other hand, Lee in  introducd Liu in  introducd Liu in , 10, Maod Liu in  introducC-approximating posets to the concept of countably QC-approximating posets. With the countably QC-approximating property, we present some characterizations of GCD lattices and generalized countably approximating posets.In this paper, we generalize the concept of countably L, ≤) be a poset. Then L with the dual order is also a poset and denoted by Lop. A principal ideal  is a set of the form ↓x = {y ∈ L∣y ≤ x} . For X⊆L, we write ↓X = {y ∈ L∣∃   x ∈ X, y ≤ x}, ↑X = {y ∈ L∣∃   x ∈ X, x ≤ y}. A subset X is a(n) lower set  if X =  ↓X . The supremum of X is denoted by ∨X or supX. A subset D of L is directed if every finite subset of D has an upper bound in D. A subset D of L is countably directed if every countable subset of D has an upper bound in D. Clearly every (countably) directed set is nonempty, and every countably directed set is directed but not vice versa. A poset L is a directed complete partially ordered set  if every directed subset of L has a supremum. A poset is said to have countably directed joins if every countably directed subset has a supremum.We quickly recall some basic notions and results . Let  to denote the power set of X and Pfin(X) to denote the set of all nonempty finite subsets of X. For a poset L, define a preorder ≤  on P(L)∖{∅} by G ≤ H if and only if ↑H⊆↑G for all G, H⊆L. That is, G ≤ H if and only if for each y ∈ H there is an element x ∈ G with x ≤ y. We say that a nonempty family F of subsets of L is (countably) directed if it is (countably) directed in the Smyth preorder. More precisely, F is directed if for all F1, F2 ∈ F, there exists F ∈ F such that F1, F2 ≤ F; that is, F⊆↑F1∩↑F2.For a set c on points of L to the nonempty subsets of L, one obtains the concept of weakly generalized countably approximating posets.Generalizing the relation ≪L be a poset having countably directed joins. A binary relation ≪c on P(L)∖{∅} is defined as follows. A≪cB if and only if for any countably directed set D⊆L, ∨  D ∈ ↑B implies D ∩↑A ≠ ∅. We write F≪cx for F≪c{x} and y≪cH for {y}≪cH. If for each x ∈ L, ↑x = ∩{↑F∣F ∈ ω(x)}, where ω(x) = {F∣F ∈ Pfin(L) and F≪cx}, then L is called a weakly generalized countably approximating poset. A weakly generalized countably approximating poset which is also a complete lattice is called a weakly generalized countably approximating lattice.Let L with the condition that for each x ∈ L, ω(x) is countably directed is called a generalized countably approximating poset (lattice) in [A weakly generalized countably approximating poset (lattice) tice) in .As a generalization of completely distributive lattice, the following concept of GCD lattices was introduced in .L be a poset. A binary relation ⊲ on P(L) is defined as follows. A⊲B if and only if whenever S is a subset of L for which ∨S exists, ∨S ∈ ↑B implies S ∩↑A ≠ ∅. A complete lattice L is called a generalized completely distributive lattice or shortly a GCD lattice, if and only if for all x ∈ L, ↑x = ∩{↑F∣F ∈ Pfin(L) and F⊲x}.Let U of a poset L is Scott-open if ↑U = U and for any directed set D⊆L, supD ∈ U implies U∩D ≠ ∅. All the Scott-open sets of L form a topology, called the Scott topology and denoted by σ(L). The complement of a Scott-open set is called a Scott-closed set. The collection of all Scott-closed sets of L is denoted by σ(L)op. The topology on L generated by {L∖↓x∣x ∈ L} as a subbase is called the upper topology and denoted by ν(L).A subset σ-Scott-open sets.Replacing directed sets with countably directed sets in L be a poset. A subset U of L is called σ-Scott-open if ↑U = U and for any countably directed set D⊆L, supD ∈ U implies U∩D ≠ ∅. All the σ-Scott-open sets of L form a topology, called the σ-Scott topology and denoted by σc(L). The complement of a σ-Scott-open set is called a σ-Scott-closed set. The collection of all σ-Scott-closed sets of L is denoted by σc(L)op.Let L, the σ-Scott topology σc(L) is closed under countably intersections and the Scott topology σ(L) is coarser than σc(L); that is, σ(L)⊆σc(L).(1) For a poset σ-Scott-closed if and only if it is a lower set and closed under countably directed joins.(2) A subset of a poset is σ-Scott-closed subsets for a poset, Mao and Xu in [C-approximating posets.To study the order structure of the lattice of all nd Xu in  introducL be a poset and x, y ∈ L. We say that x is σ-beneath y, denoted by x≺σy, if for any nonempty σ-Scott-closed set F⊆L for which ∨F exists, ∨F ≥ y always implies that x ∈ F. Poset L is said to be countably C-approximating if for each x ∈ L, x = ∨↓σ≺x, where ↓σ≺x = {y ∈ L∣y≺σx}. A complete lattice which is also countably C-approximating is called a countably C-approximating lattice.Let L, the lattice σc(L)op is countably C-approximating.For a poset L be a poset and C ∈ σc(σc(L)op)op. It is straightforward to check that ⋁σc(L)opC = ∪C. For each F ∈ σc(L)op, we have that F = ⋁σc(L)op{↓x∣x ∈ F}. Suppose C ∈ σc(σc(L)op)op with ⋁σc(L)opC⊇F. Then for each x ∈ F, since ⋁σc(L)opC = ∪C⊇F, there exists A ∈ C such that x ∈ A. Noticing that A ∈ σc(L)op is a lower set, we have ↓x⊆A ∈ C. It follows from C ∈ σc(σc(L)op)op being a lower set that ↓x ∈ C. Thus by x≺σF holds in σc(L)op. Hence, F = ⋁σc(L)op{↓x∣x ∈ F}⊆∨↓σ≺F⊆F. So, F = ∨↓σ≺F and by the arbitrariness of F ∈ σc(L)op, we conclude that σc(L)op is countably C-approximating.Let QC-approximating posets. Firstly, we generalize the relation ≺σ on points of a poset L to the nonempty subsets of L.In this section, we introduce the concept of countably L, the σ-beneath relation ≺σ on nonempty subsets of L is defined as follows: A≺σB if and only if whenever S is a nonempty σ-Scott-closed subset of L for which ∨S exists, ∨S ∈ ↑B implies S ∩↑A ≠ ∅. We write F≺σx for F≺σ{x}. Set c(x) = {F∣F ∈ Pfin(L)  and  F≺σx}.For a poset The next proposition is basic and the proof is omitted.L be a poset. ThenG, H⊆L, G≺σH⇒G ≤ H;∀G, H⊆L, G≺σH⇔∀h ∈ H, G≺σh;∀E, F, G, H⊆L, E ≤ G≺σH ≤ F⇒E≺σF;∀x, y ∈ L, {x}≺σ{y}⇔x≺σy.∀Let σ, we have the concept of countably QC-approximating posets.With the relation ≺L is said to be countably quasi-C-approximating, shortly countably QC-approximating, if for all x ∈ L, ↑x = ∩{↑F∣F ∈ c(x)}. A countably QC-approximating poset which is also a complete lattice is called a countably QC-approximating lattice.A poset C-approximating posets are countably QC-approximating.Countably L be a countably C-approximating poset. Then for all x ∈ L,F∣F ∈ c(x)} = ↑x. By L is countably QC-approximating.Let By L, the lattice σc(L)op is countably QC-approximating.For any poset QC-approximating lattices and GCD lattices.In the sequel, we explore relationships between countably Every GCD lattice is weakly generalized countably approximating.L be a GCD lattice. For all x ∈ L and F ∈ Pfin(L), F⊲x implies F≪cx. Then ↑x⊆∩{↑F∣F ∈ ω(x)}⊆∩{↑F∣F ∈ Pfin(L) and F⊲x} = ↑x. So ↑x = ∩{↑F∣F ∈ ω(x)}. By L is weakly generalized countably approximating.Let QC-approximating.Every GCD lattice is countably L be a GCD lattice. For each x ∈ L and F ∈ Pfin(L), F⊲x implies F≺σx. Then ↑x⊆∩{↑F∣F ∈ c(x)}⊆∩{↑F∣F ∈ Pfin(L) and F⊲x} = ↑x. Thus ↑x = ∩{↑F∣F ∈ c(x)}. By L is countably QC-approximating.Let The following theorem characterizes GCD lattices.L be a complete lattice. Then the following statements are equivalent:L is a GCD lattice;L is countably QC-approximating and weakly generalized countably approximating.Let (1)⇒(2): follows from Propositions L is countably QC-approximating and weakly generalized countably approximating. Then for each x ∈ L, by the weakly generalized countably approximating property of L, we have ↑x = ∩{↑F∣F ∈ ω(x)}. Now for each F ∈ ω(x), we show that ↑F = ∩{↑F′∣F′ ∈ Pfin(L) and F′≺σF}. To this end, it suffices to show that ∩{↑F′∣F′ ∈ Pfin(L) and F′≺σF}⊆↑F. Suppose t ∈ ∩{↑F′∣F′ ∈ Pfin(L) and F′≺σF} and t ∉ ↑F. Then for any yF ∈ F, t ∉ ↑yF. By the countably QC-approximating property of L, there exists FyF ∈ c(yF) such that FyF≺σyF and t ∉ ↑FyF. Let t ∈ ∩{↑F′∣F′ ∈ Pfin(L) and F′≺σF}. Thus ↑x = ∩{↑F∣F ∈ ω(x)} = ∩{↑F′∣F′ ∈ Pfin(L), ∃F ∈ Pfin(L) such that F′≺σF≪cx}.(2)⇒(1): suppose that F′≺σF≪cx, we will show that F′⊲x. For any A⊆L with ∨A ≥ x, let G = {∨E∣E is a countable subset of A}. Then G is a countably directed set and ∨G = ∨A ∈ ↑x. Since F≪cx, there exists a countable subset E⊆A such that ∨E = ∨↓E ∈ ↑F. By E is σ-Scott-closed. It follows from F′≺σF that ↓E∩↑F′ ≠ ∅. This implies A∩↑F′ ≠ ∅, showing that F′⊲x. Thus, ↑x⊆∩{↑W∣W ∈ Pfin(L), W⊲x}⊆∩{↑F′∣F′ ∈ Pfin(L), ∃ F ∈ Pfin(L), F′≺σF≪cx} = ↑x. So, ↑x = ∩{↑W∣W ∈ Pfin(L) and W⊲x}. Therefore, L is a GCD lattice.Suppose L is called a hypercontinuous poset x} is directed and x = sup{y ∈ L∣y≺ν(L)x}, where y≺ν(L)x⇔x ∈ intν(L)↑y. A hypercontinuous poset which is also a complete lattice is called a hypercontinuous lattice.Recall that a poset set (see ) if for L be a complete lattice. Then L is a GCD lattice if and only if Lop is a hypercontinuous lattice.Let L, both L and Lop are continuous, and ν(L) = σ(L). It follows from  is hypercontinuous.Let σc(L)op is a GCD lattice. The following theorem gives comprehensive characterizations of generalized countably approximating posets.So, in view of L be a poset having countably directed joins. Then the following statements are equivalent:L is a generalized countably approximating poset;σc(L) is a hypercontinuous lattice;σc(L)op is a GCD lattice;σc(L)op is a weakly generalized countably approximating lattice.Let (i)⇔(ii) by (ii)⇔(iii) by (iii)⇔(iv) follows from


The objective of the current investigation was to explore whether upper body accelerations obtained during gait provide sensitive measures of postural control in people with Parkinson's disease (PD). Thirteen people with PD (70 ± 11 years) and nineteen age-matched controls (70 ± 7 years) walked continuously for two minutes while wearing three inertial sensors located on their lower back (L5), shoulder level (C7), and head. Magnitude (root mean square (RMS)), attenuation (attenuation coefficient), and smoothness  of the accelerations were calculated. People with PD demonstrated greater RMS, particularly in the mediolateral direction, but similar harmonic ratio of head accelerations compared to controls. In addition, they did not attenuate accelerations through the trunk and neck as well as control participants. Our findings indicate that measuring upper body movement provides unique information regarding postural control in PD and that poor attenuation of acceleration from the pelvis to the head contributes to impaired head control. This information is simple to measure and appears to be sensitive to PD and, consequently, is proposed to benefit researchers and clinicians. People with Parkinson's disease (PD) walk with a gait pattern that is characterised by slowness (bradykinesia), muscle rigidity, and postural instability –3. As thThe recent development of small and inexpensive wireless inertial sensors has helped facilitate routine measurement of postural control during gait in the clinic, laboratory, and the community. Emerging evidence suggests that measuring upper body acceleration during gait using inertial sensors can objectively quantify differences in gait patterns between those with and without PD , 10. It Despite emerging evidence that maintaining head stability during gait is a key determinant of postural control –16, acceThe objective of the current investigation was to explore whether upper body accelerations obtained during gait provide sensitive proxy measures of postural control in people with Parkinson's disease (PD). More specifically, the aims of this study were to assess the magnitude, attenuation, and smoothness of upper body accelerations in people with PD compared to age-matched controls. We tested the hypotheses that: people with PD would demonstrate impaired smoothness and attenuation of accelerations. To address these aims, accelerations of the head, trunk, and pelvis were assessed during gait in a cohort of people with PD and an age-matched control group.A subsection of community dwelling older adults and people with PD were tested as part of the ongoing ICICLE-PD  Gait study , 23. ParAll participants walked for two minutes at their preferred pace around a 25 m circuit, fully described in . SpatiotTo ensure only steady-state, straight-line walking was analysed, only the portion of the acceleration data recorded while participants who were in contact with the Gaitrite walkway was used. As detailed in Mazzà et al. , prior tThe magnitude of accelerations was calculated using the root mean square (RMS) of the accelerations, measured by each sensor for each stride in the Anteroposterior (AP), Mediolateral (ML), and Vertical (V) directions.The ability to attenuate accelerations through the upper body was quantified using the attenuation coefficient. The attenuation coefficient describes the ability to reduce accelerations from inferior to the superior anatomical locations and was calculated using the RMS values for each stride.H), shoulder (RMSS), and pelvis (RMSP) as follows [CPH representing the attenuation from the pelvis to the head, CPS representing the attenuation from the pelvis to the shoulder, and CSH representing the attenuation from the shoulder to the head. Each equation provides a percentage representing the amount of acceleration that is attenuated from the inferior sensors to the superiorly located sensor. A positive coefficient indicates reduced acceleration at the superiorly located sensor relative to the inferiorly located sensor. A negative coefficient value indicates a greater acceleration at the superiorly located sensor.The attenuation coefficients were computed using the RMS values of the head (RMS follows –20: (1)CWe quantified the smoothness of upper body accelerations using the harmonic ratio (HR). The HR accurately describes the step-to-step symmetry within a stride but for upper body gait analysis is also commonly referred to as a measure smoothness . The HR For the AP and V Components, the HR Was Defined as:For the ML Component, the HR Was Defined as:vourable , 31. Folvourable , 14.t-tests were used to test the difference between groups for the magnitude, attenuation, and smoothness of accelerations. The level of significance was set at P = 0.05. Given the exploratory nature of this study, the P value was not adjusted for multiple comparisons.A series of two-tailed paired The characteristics of the participants are reported in 2 versus 0.86 ± 0.21 m/s2, P = 0.024) but not at the pelvis or the shoulder level. There were no other significant between-group differences although AP and V head accelerations tended to be greater in the PD group . For CPS, a significant difference existed between PD and controls in the AP , as well as the ML direction . For CSH, a significant difference existed between the PD and the control group in the ML direction .People with PD did not attenuate AP or ML accelerations as well as controls . For CPThe HRs normalised to gait speed showed no significant differences between the PD and control participants .Our current investigation provides evidence that upper body accelerations obtained during gait provide sensitive measures of postural control in people with Parkinson's disease (PD). As hypothesised, the results of this study showed that people with PD walked with altered upper body accelerations compared to age-matched controls. In particular, people with PD walked with greater magnitude of ML head accelerations and demonstrated impaired attenuation of accelerations from the pelvis and neck to the head. In contrast to our hypothesis, smoothness of upper body accelerations as measured by the HR was not significantly affected in this sample of PD.To our knowledge, this is the first study to show impaired head stability in people with PD using inertial sensors. A greater magnitude of ML head acceleration was found for the PD group. This was interpreted as a result of poor postural control for the PD participants and a failure to stabilise their head in space , 21. Hig en bloc movement and axial rigidity. It has previously been stated that increased rigidity may cause underlying changes in the physiological and mechanical functioning of the axial muscles which results in en bloc movement, where the head, trunk and pelvis move together as one rigid unit [Alternatively, being computed as a ratio between accelerations measured during the same trial , the coegid unit , 32. It Interestingly, the findings regarding attenuation coefficients were strongest in the ML direction. Similar results were found even when analysing healthy elderly subjects . The facP = 0.106). Further research is required to determine the effectiveness of harmonic ratios as a sensitive measure to PD at different stages of their disease progression, as well as its ability to predict future falls.In contrast with our hypothesis and previous studies , the smoClinicians require objective measures to assess postural control during locomotion in people with PD to supplement standard clinical assessments and conventional rating scales which are not sensitive to subtle postural control disturbance , 33, 34.The current investigation suggests that assessing upper body acceleration offers additional and unique information about postural control during gait in people with PD. In particular, the magnitude of ML head accelerations and attenuation of upper body acceleration appear sensitive to PD and consequently hold promise as useful proxy measures that can be utilised in clinical and community settings.


Osteogenesis imperfecta (OI) is an inherited bone and connective tissue disorder associated with the lifelong occurrence of frequent fractures following even mild trauma. Hearing loss is frequently reported in patients with OI. Objective: to examine the ratio of hearing loss in children with OI, and the relationship between audiological findings and CT images of temporal bone in children with OI. Subject and methods: forty – two children aged 5 to 17 years with OI were included in the study. The patients have type A of tympano and were mesured thresold of hearing by play audiometry. CT imaging was performed in 8 cases as well. Imaging abnormalities were correlated with clinical phenotypes and severity of hearing loss deduced from audiograms. Results: Hearing loss of all etiologies was observed in 28.05 % of ears in studied OI patients. Sensorineural and mixed hearing loss was observed in 4.88% and conductive hearing loss was detected in 23.17% of ears. CT revealed bone – bridge image in the middle ear (10/16 ears), hypodense foci in the fissula ante fenestram (4/16 ears) and cochlear (2/16 ears), abnormal stape ( 5/16 ears). Conclusions: hearing loss in children with osteogenesis imperfecta is quite frequent. We have all type of hearing loss, but the conductive of hearing loss have highest ratio. The site of abnormal on temporal bone CT images in OI corresponds to presence and type of hearing loss determined by audiometry.


Here we report the presence of ectopic retina-like tissue in some of the transgenic mice, young and aged. The ectopic retina-like tissue contained cells positive for photoreceptor proteins Crx, recoverin, red opsin, and rhodopsin, and cells positive for proteins that label other types of retinal neurons, including AP2α and Pax6 for amacrine cells, Otx2 for bipolar cells, and Brn3A for ganglion cells. The retina-like tissue often co-existed with darkly pigmented tissue positive for RPE proteins: cytokeratin 18, Otx2, and RPE65. The ectopic retina-like tissue was detected in the subretinal space, including two retinae co-existing in the same eye, and/or in the optic nerve or in the vicinity of the optic nerve head. On rare occasions, it was detected in the choroid and in the vicinity of the ciliary body. The presence of ectopic retina-like tissue in the transgenic mouse supports the possibility of inducing retinal regeneration in the mammalian eyes through gene-directed reprograming.Degeneration of retinal neurons is an underlying cause of several major types of blinding diseases, and effective therapies remain to be developed. The suppositive strategy of repopulating a degenerative retina with new cells generated onsite faces serious challenges, because the mammalian retina seems to lack the ability to regenerate itself or replace its lost neurons. We investigated the possibility of using a transcriptional factor with proneural activities to reprogram ocular tissue with regenerative capability to give rise to retinal cells. Transgenic mice were generated with DNA constructs that targeted the expression in the retinal pigment epithelium of proneural gene  The neural retina is a thin tissue in the back of the eye and is responsible to initiate vision when the light enters the eye. In the retina, photoreceptor cells capture photons and generate electrophysiological signals. The signals are modulated by interneurons comprised of horizontal, bipolar, and amacrine cells and relayed by retinal ganglion cells to the brain for visual perception. Immediately outside the neural retina lies a single-layered, darkly pigmented, non-neural tissue—the retinal pigment epithelium (RPE), which plays important roles in retinal physiology. Retinal degeneration and RPE atrophy are the main causes of many common forms of visual loss, including retinitis pigmentosa, age-related macular degeneration, diabetic retinopathy, and glaucoma. The low quality of life of people with visual loss has spurred research interests in a wide range of potential therapeutic approaches, including retinal regeneration.Naturally occurring regeneration from retinal stem cells would be ideal for producing new cells to repopulate a retina inflicted with degeneration. However, the presence of retinal stem cells in adult mammals remains elusive ,2. The rOne of the ocular tissues being investigated is the RPE. Anatomically, the RPE would be a convenient source of new cells to repopulate a degenerative retina. Developmentally, the RPE and the retina share a common origin, the optic vesicle. Biologically, mammalian RPE, including that of human, is capable of two cellular events: proliferation and plasticity. It has been shown that in a mature eye a small population in the periphery RPE maintain mitotic activities . The majNeurogenin1 (ngn1) and ngn3, members of the neurogenin subfamily of the basic Helix-Loop-Helix family of transcription factors and homologous to the Drosophila proneural gene atonal, can efficiently reprogram differentiated RPE cells of embryonic chick to develop into cells displaying molecular, morphological, and physiological properties of young photoreceptor cells, with small fractions of the product cells expressing markers of other types of retinal neurons [ngn1 from the promoter of Bestrophin1 (PVMD2-ngn1) or ngn3 from the promoter of RPE65 (PRPE65-ngn3), photoreceptor-like cells were detected in the subretinal space in both young and aged animals [A gene-directed reprograming of differentiated RPE to serve as a potential source of new retinal neurons has been tested with chick and mammalian cells.  neurons . Similar neurons . In tran animals .eyeless (Pax6) produces ectopic eyes on the wings, the legs, and the antennae [Pax6 [Six3 [Six6 [Pax6 and Six6) and medaka fish (Six3). Ectopic expression in the hindbrain-midbrain area of Six3 from Pax2 promoter leads to the formation of ectopic optic vesicle-like structures in about ½ of the transgenic mice [An even more audacious and elusive aim is to regenerate a retina in the mammalian eye. In Drosophila, targeted misexpression of antennae . This haae [Pax6 , Six3 [3x6 [Six3 , and Sixx3 [Six6  each cannic mice . WhetherVMD2-ngn1 and PRPE65-ngn3 designs in generating transgenic mice [To determine whether the Pnic mice  could pr2 inhalation and cervical dislocation .All experimental procedures involving the use of animals adhered to the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health and were approved by the Institutional Animal Care and Use Committee of the University of Alabama at Birmingham (Permit Number: 140909197). All efforts were made to minimize suffering. Euthanization was carried out with COBestrophin1 promoter (PVMD2) and the sequence (−737 to 0) of mouse RPE65 promoter (PRPE65) were PCR amplified, cloned into pGEM-T, and sequence verified. PRPE65 (or PVMD2) was inserted into vector pCI replacing PCMV to drive transgene expression in the RPE. The coding sequences of human ngn1 and ngn3 were RT-PCR amplified and cloned into pGEM-T. After sequence verification, the coding regions were linked with PVMD2 and PRPE65 to generate pCI-PVDM2-ngn1 and pCI-PRPE65-ngn3, respectively, which were used to create potential founders at the University of Alabama at Birmingham Transgenic Facility. Transgenic animals were identified by PCR genotyping with DNA from tail snips.Transgenic mice were generated as previously described . BrieflyTissue preparation, cryosectioning, and immunohistochemistry were performed as previously described . BrieflyVMD2-ngn1 and PRPE65-ngn3 transgenic mice [ngn1 and ngn3 to channel RPE’s proliferation and plasticity towards de novo generation of retinal neurons, particularly photoreceptor cells, as observed with cultured RPE cells derived from chick embryos [Bestrophin1 and RPE65 were used because they are primarily, although not exclusively, expressed in differentiated RPE cells [ngn1 and ngn3 expression during neurogenesis observed in the chick retina. In approximately 60% of offspring from ~50% of founders, there are photoreceptor-like cells in the subretinal space [The underlying theme of the creation of Pnic mice  was to u embryos . The proPE cells ,40. The al space . The reaVMD2-ngn1 and PRPE65-ngn3 mice of extra piece of tissue with histological nuclear demarcation typical of the retina. In the eye of a 2-week-old PRPE65-ngn3 mouse, a small piece of such tissue was detected at the posterior region . We estimated that ~10% of the transgenic mice displaying photoreceptor-like cells in the subretinal space [Eyes of adult mice were then analyzed to examine whether they might contain retina-like tissue, regardless of the tissue either having survived into adulthood or being generated in adulthood. In a 2-month-old Psame eye . Each ofsame eye . The ONLsame eye , and thesame eye , Otx2  expressed in Müller glia, cellular retinaldehyde binding protein (CRALBP) expressed in Müller glia (as well as in the RPE), and glial fibrillary acidic protein (GFAP) expressed in astrocytes and activated Müller glia, and antibody A10-33/1 against endothelial cells of blood vessels. In eyes with a relatively small amount of photoreceptor-like cells present in the subretinal space, CRALBPtributed , and so al cells . Abnormaal cells . In thisd region . In thisd region  conspicud region , an indime scant .RPE65-ngn3 mouse, DAPI staining showed a piece of densely populated tissue, a histological mark of the retina, in the optic nerve that is otherwise loosely populated . It could also be a result of cell death from inflammation, reactive gliosis, lacking trophic supports normally derived from postsynaptic targets but absent for the retinal-like tissue, and lacking oxygen supply to maintain all retinal layers due to diminished blood vessels. Like in the retina, cells in the ONL-like layer expressed Crx, recoverin, red opsin, and rhodopsin, cells in the INL-like layer were positive for Otx2, AP2α, and Pax6, and cells in the GCL-like layer were positive for Brn3A. These data suggest that retina-like tissue was induced by ectopic ngn1 expression from the Bestrophin1 promoter and by ectopic ngn3 expression from the RPE65 promoter.Retinal regeneration in the mammalian eye remains an elusive goal. In these experiments we showed the presence in PAnatomically, the retina-like tissue localized both within and outside the normal locale of the retina. There was substantial occurrence of retinal-like tissue present in the optic nerve region. It is possible that these retina-like tissue had originated from the retina and had migrated and/or extended into the optic nerve region. An alternative scenario is that it was produced from the optic nerve region, particularly the optic head region.Rather remarkably, retina-like tissue at an ectopic location was detected in young and adult mice. In the adult animal, the tissue might be generated at a young age, including an embryonic stage, and survived well into adulthood. Alternatively, it was generated during adulthood. Either scenario is intriguing and warrants corroboration with further investigation. Perhaps equally interesting and important, the retina-like tissue was often associated with RPE-like tissue. Considering RPE’s importance in retina’s function and health, the coexistence of retina-like and RPE-like tissues enhances the possibility of exploring this reprogramming strategy in inducing functional, retinal regeneration in the mammalian eye. Keep in mind, however, functional regeneration also requires successful establishment of blood supply to the regenerated retina and integration of the new one into the existing visual circuitry.ngn1 or ngn3 and the cells that expressed the transgene under PVMD2 or PRPE65; neither is well understood. The selection of ngn1 and ngn3 was based on previous studies with the chick system. Ngn1 and ngn3 are found to be the frontrunners among more than 20 genes and factors tested in terms of efficiently reprogramming RPE cells to differentiate into cells displaying photoreceptor traits [ngn1 and ngn3 are transiently expressed in early developmental stages, when active cell proliferation and cell fate specification take place [ngn3 promotes early retinal neurogenesis, and ngn1 is one of the downstream genes of ngn3 [ngn1 and ngn3 participate in retinal neurogenesis in the mouse retina.Intriguing as they may seem, our data raise more questions than they answered. The two key ingredients in this study were proneural gene r traits . In the ke place ,44. Gain of ngn3 . CurrentVMD2-ngn1 and PRPE65-ngn3 DNA constructs, which lacked additional temporal and spatial regulatory elements for a tighter regulation of the transgene. Thus, the levels, the locales, and time of the transgene expression might have varied substantially. In addition, current information does not rule out the expression of Bestrophin1 and RPE65 outside the RPE, including in the developing retina and in Müller glia [Despite of the design that aimed at targeting the transgene expression in differentiated RPE cells, the RPE might not be the source, at least not the sole source, of the ectopic retinal-like tissue. The transgenic mice analyzed in this study were created with the simple Pler glia , which hVMD2-ngn1 and PRPE65-ngn3 transgenic mice, and eyecup explants of “sclera+choroid+RPE” from adult PVMD2-ngn1 and PRPE65-ngn3 animals can generate, de novo, photoreceptor-like cells in culture [The scenario of RPE as one, although not necessarily the only, source of the retina-like tissue is consistent with published data. Stem-like cells with multi-potentials have been isolated from the RPE of human adults . Cells s culture , providiWhen located in the subretinal space, the retinal-like tissue might displayed a reversed tissue polarity in regarding to the RPE, as observed in the classic RPE-to-retina transdifferentiation. However, unlike the classic RPE-to-retina transdifferentiation where the new retina is produced at the expense of the RPE, the RPE remained in our transgenic mice, as previously shown . Furtherngn1 and ngn3 expression from PVMD2 and PRPE65 and had given rise to the new tissues. No information is available on the identities of these cells and whether they intrinsically remained multi-potential progenitor properties or such properties were bestowed by ectopic expression of ngn1 and ngn3. In-depth investigation with a wide scope is needed for a better understanding of the process before applying the strategy for retinal regeneration in the mammalian eye.The presence of retina-like tissue at ectopic locations, particularly in the optic nerve, the vicinity of optic nerve head, the choroid, and exterior to the eyecup and the co-presence of RPE-like tissue with retina-like tissue imply the presence of progenitor-like cells, including those embedded inside the RPE. These progenitor-like cells had responded to


Osteoporosis is a disease that weakens bones and is known to be “silent,” but it does not mean that it should be ignored. According to some researches, osteoporosis causes more than 8.9 million fractures every year, resulting in an osteoporotic fracture every 3 s and by 2050 the world-wide occurrence of hip fracture is estimated to increase by 310% in men and 240% in women . Mainly bones consist of minerals such as a matrix of hydroxyapatite  and other minerals which are embedded in a cross-linked collagen matrix. Normally, there is a delicate balance between bone formation and bone re-absorption, which if bone re-absorption becomes dominant osteopenia and osteoporosis may occur Larsen, . In anotOn the other hand, according to some researches, it has been shown that there is a correlation between BMD loss in osteoporotic patients and conductive hearing loss; in those patients over a specific age, changing the structure of the ossicles or hormonal mechanism in hearing may correlate BMD loss with hearing loss Shafer, .In fact, the hearing and balance systems are connected inside the inner ear and hearing loss can affect balance system. Basically, Balance is a complex that receives sensory information from a variety of organs  and integrates it to inform the body of where it is. Furthermore, information from the vestibular system of the inner ear  is sent to the brain. Semicircular canals in the inner ear are able to sense changes in movement of the body. Due to these changes, endolymph within the canals moves inner ear’s hair cells. As a result, position of the head is sensed by hair cells. Normally, the information coming from the ears perfectly matches the information coming from the eyes and the sensors in the joints and balance complex completes (Standring, In conclusion, we hypothesize that in osteoporotic patients, who have low BMD, hearing loss occurs and as a result balance problems happen. In spite of the fact that some researchers have shown that osteoporotic patients have balance disorders and hearing loss (Henkin et al.,


Traumatic pneumolabyrinth is a relatively rare entity. We report the case of a unilaterally deaf woman with pneumolabyrinth who had suffered penetrating injury 15 years ago. This past history indicated that the case was late pneumolabyrinth occurring from undiagnosed old posttraumatic perilymphatic fistula. In Japan, most cases of traumatic pneumolabyrinth are caused by penetrating injury with an ear pick. Dizziness often improves within several months. Immediate surgical intervention is recommended for hearing loss, but the hearing outcome is not satisfactory. An appropriate strategy should be selected based on the interval to surgery, bone conduction hearing level at disease onset, stapes lesions, and location of air. Pneumolabyrinth is a condition caused by air in the labyrinth, which indicates that a perilymphatic fistula (PLF) is present in the affected ear. However, patients with traumatic PLF seldom develop pneumolabyrinth. Here, we report a case of pneumolabyrinth that may have been caused by a penetrating injury of the tympanic membrane that occurred 15 years before presentation. This interesting case suggests that pneumolabyrinth may be among the differential diagnoses for recurrent dizziness.A 25-year-old woman was admitted to hospital with a complaint of longstanding right-sided profound hearing loss. Her hearing impairment had begun 15 years ago. At that time, she suffered traumatic tympanic membrane perforation due to penetration by an ear pick. She had sudden profound hearing loss at the onset of injury and it was unclear that she had felt vertigo after injury. Conservative therapy was performed at ENT clinic nearby. Although perforation of tympanic membrane was healed completely, profound hearing loss had remained. She mentioned the degree of hearing loss had not changed. She had felt lightheadedness about 7 years ago.An otoscopic examination revealed no perforation in her right ear, but pure tone audiometry showed right profound hearing loss . ImagingTwo years later, she was admitted again for follow-up of hearing loss. She sometimes had dizziness or lightheadedness. Pure tone audiometry showed right profound hearing loss similar to before . EquilibThe number of reports on pneumolabyrinth is smaller than those on PLF but has shown a slight increase due to improved accuracy of imaging modalities. Hidaka et al.  suggesteAccording to medical interview, profound hearing loss occurred immediately after injury and was supposed to be permanent. Intrusion of stapes found in CBCT image indicated the possibility of old posttraumatic PLF. Symptoms associated with PLF tend to appear rapidly and the degree of hearing loss varies from mild to profound. Subacute deterioration of hearing level has been documented in several reports –6 and alThe unique aspect of this case is the long interval between injury and diagnosis of pneumolabyrinth by CBCT. It was impossible to make sure that pneumolabyrinth had occurred soon after injury. Intrusion of stapes suggested that there had been posttraumatic PLF. There was no causative event other than penetrating injury by ear pick 15 years ago. Air that enters the inner ear usually disappears within several weeks. Pneumolabyrinth detected by CBCT suggested that air was trapped recently and that PLF did not heal completely. In the patient with PLF, elevated atmospheric pressure in the middle ear cavity may induce fainting and dizziness. If excessive elevation of the middle ear pressure happens, entrapment of air in the vestibule may occur. Nurre et al.  describeTreatment of dizziness has a good prognosis, but treatment of hearing loss is difficult and often unsuccessful. Most reports have shown unsatisfactory hearing outcomes regardless of the treatment strategy. Hidaka et al.  suggesteTsubota et al.  found thHidaka et al.  also fouPrevention from worsening of hearing loss is important. Achache et al.  emphasizPneumolabyrinth may occur in case PLF is present, even if the interval between diagnosis and causative event is very long. An appropriate decision on the treatment strategy requires consideration of the timing of surgery, the bone conduction level at onset of disease, stapes lesions, and location of air in the inner ear.


Mycoplasma meleagridis and Mycoplasma gallinarum are bacteria that affect birds, but little is known about the genetic basis of their interaction with chickens and other poultry. Here, we sequenced the genomes of M. meleagridis strain MM_26B8_IPT and M. gallinarum strain Mgn_IPT, both isolated from chickens showing respiratory symptoms, poor growth, reduction in hatchability, and loss of production. Mycoplasma meleagridis and Mycoplasma gallinarum belong to the hominis phylogenetic group within the Mollicutes class. Turkey was considered to be, for a long time, the specific host for M. meleagridis .eagridis , until ieagridis . The ubitigators . It is cCC 17529 , M. meleM. meleagridis and M. gallinarum, we sequenced the genomes of two field strains of M. meleagridis MM_26B8_IPT and M. gallinarum Mgn_IPT.To facilitate our understanding of the host tropism and pathogenic potential of Both strains were isolated from chicken farms located in the Cap Bon region (northeast) of Tunisia. Strains were cultivated in Frey’s medium  and puriThe MM_26B8_IPT and Mgn_IPT genomes were sequenced, annotated, and analyzed as described previously . BrieflyThe MM_26B8_IPT genome has a size of 658,083 bp (assembled in 32 contigs), whereas the Mgn_IPT one is smaller, consisting of 800,663 bp (organized in 56 contigs). The G+C content in the two genomes was almost the same, around 26%. A total of 539 and 601 coding sequences (CDSs) were identified in MM_26B8_IPT and Mgn_IPT, respectively. The more the genome was reduced, the greater was the coding density (90.44% in MM_26B8_IPT versus 87.40% in Mgn_IPT). Both strains were found to contain 33 tRNAs and a single copy of tmRNA.Among the 9 and 10 genes involved in the stress response determined in MM_26B8_IPT and Mgn_IPT, respectively, 8 heat shock proteins  were shared. In addition, 17 and 14 genes potentially implicated in virulence and resistance to antibiotics and toxic compounds were identified in MM_26B8_IPT and Mgn_IPT, respectively.M. meleagridis and M. gallinarum, isolated from Tunisian chickens will help further the study of their host specificity and pathogenic potential.Further analyses of the genome sequences of these two avian field strains, M. meleagridis and M. gallinarum field strains have been deposited at DDBJ/EMBL/GenBank under the accession numbers LVWO00000000 and LVLH00000000, respectively. The versions described in this paper are versions LVWO00000001 for M. meleagridis MM_26B8_IPT and LVLH00000001 for M. gallinarum Mgn_IPT.These whole-genome shotgun projects of


Endocrine disrupting chemicals (EDCs) are chemicals that have the capacity to interfere with normal endocrine systems. Two EDCs, bisphenol A (BPA) and triclosan (TCS), are mass-produced and widespread. They both have estrogenic properties and similar chemical structures and pharmacokinetic features and have been detected in human fluids and tissues. Clinical evidence has suggested a positive association between BPA exposure and implantation failure in IVF patients. Studies in mouse models have suggested that preimplantation exposure to BPA and TCS can lead to implantation failure. This paper reviews the relationship between preimplantation exposure to BPA and TCS and implantation failure and discusses the remaining problems and possible solutions. During the last few decades, the incidence of human infertility has significantly increased in many countries, such as the United States and China –5. The rβ-estradiol [Some chemicals which have been widely used for decades have recently been found to have the ability to disrupt endocrine function in humans. They are called endocrine disrupting chemicals (EDCs). Bisphenol A (BPA) and triclosan (TCS) are EDCs with similar chemical structures to 17stradiol  . Women with urinary BPA above the lowest quartile had decreased blastocyst formation (trend test P = 0.08). Ehrlich et al. [In 2005, Sugiura-Ogasawara et al.  reportedh et al.  also claThe correlation between the concentration of TCS in pregnant women and female infertility remains unclear due to a lack of investigation. However, the similarity of the distribution, chemical structures, and estrogenic activity of BPA and TCS suggests the possible involvement of TCS in implantation failure.Implantation failure could be caused by the embryo itself, inadequate uterine receptivity, or defects in communication between the embryo and the endometrium. It is generally assumed that the embryo itself is probably only responsible for one-third of IVF failures, and the other two-thirds of implantation failures occur due to impaired uterine receptivity or defects in embryo-endometrium communication –60. AnimCrawford and deCatanzaro  studied Takai et al.  showed tα, ERβ, PRA, PRB, and hCG/LH-R, reduced the mRNA transcription of dPRL, and stimulated secretion of MIF in human endometrial stromal cells in vitro.Uterine receptivity and embryo development are both critical for successful implantation. Coordinated actions of progesterone and estrogen play a critical role in creating a receptive uterine environment, embryo development, and embryo migration through the oviduct , 63. EstSeveral studies have shown that BPA can impair the development of mouse embryos and that this effect can be counteracted by Tamoxifen (an estrogen receptor modulator) , 61, 68.β-estradiol have similar chemical structures and they are all fat-soluble chemicals. BPA and TCS both have estrogenic activity, share similar pharmacokinetic features, and can be detected in human fluids and tissues. Evidence from clinical studies and animal models supports the assumption that preimplantation exposure to BPA and TCS could lead to implantation failure in humans.BPA, TCS, and 17α (estrogen receptor α) and PR (progesterone receptor) in the uterus of mice [In addition, preimplantation exposure to BPA can nonmonotonically change the expression of the ER of mice . Thus it of mice  reportedBPA also can increase the luminal area and luminal cell height of the mouse uterus on gestational day 6 following subcutaneous injections of BPA on days 1–4 of gestation . These mVarayoud et al.  reportedSome studies have suggested that exposure to BPA could affect the meiotic maturation of oocytes in humans and mice –76.Crawford and deCatanzaro  suggesteIn the clinic, fertilization can only be confirmed afterwards. There is no way to detect embryo implantation failure in the clinic except for patients undergoing in vitro fertilization. The most common way to diagnose pregnancy is by testing human chorionic gonadotrophin (HCG) in urine samples. However, HCG is secreted by the syncytiotrophoblast and is detectable in maternal blood two days after the implantation of the embryo . Thus, iHowever, the most sensitive time for BPA and TCS to influence implantation remains unknown. Although the preimplantation period might be a sensitive time frame for BPA and TCS exposure, it might not be the most sensitive and important one.The procedure of in vitro fertilization and embryo transfer (IVF/ET) has provided some possible ways to identify the most dangerous time frame for TCS and BPA exposure in humans. Only in those patients would we know the exact time of fertilization, the condition of the oocyte prefertilization, and the embryo's preimplantation.Recently, Ehrlich et al. , 16 studMany studies have investigated the association between BPA exposure and clinical outcomes in IVF patients –16, 18. It is not easy to maintain low TCS and BPA exposure levels due to their widespread existence. And recently it has been reported that BPA-free plastic products also release other chemicals with estrogenic activity . Fortuna


The presence of TCs has been identified in atria, ventricles and many other tissues and organ, but not yet in heart valves. We used transmission electron microscopy and immunofluorescence methods  to provide evidence for the existence of TCs in human heart valves, including mitral valve, tricuspid valve and aortic valve. TCs are found in both apex and base of heart valves, with a similar density of 27–28 cells/mm2 in mitral valve, tricuspid valve and aortic valve. Since TCs are known for the participation in regeneration or repair biological processes, it remains to be determined how TCs contributes to the valve attempts to re-establish normal structure and function following injury, especially a complex junction was found between TCs and a putative stem (progenitor) cell.Valve interstitial cells (VICs) are responsible for maintaining the structural integrity and dynamic behaviour of the valve. Telocytes (TCs), a peculiar type of interstitial cells, have been recently identified by Popescu*s group in epicardium, myocardium and endocardium (visit  Heart valves are outgrowths from the endocardium which enable the blood to flow in a unidirectional way . The heaTwo types of cells have been identified within heart valves including endothelial cells and valve interstitial cells (VICs) . Endothehttp://www.telocytes.com). Telocytes are described as interstitial cells with extremely long and thin prolongations, called telopodes (Tps) [Telocytes (TCs), a special type of interstitial cells, have been identified by Popescu*s group –27 and tes (Tps) ,11,18. Tes (Tps) ,11,18. Aes (Tps) ,14. Tp hes (Tps) . This exes (Tps) . They rees (Tps) . The mices (Tps) ,43. Morees (Tps) –33. ThusHuman heart valves including mitral valve, tricuspid valve and aortic valve were obtained from prospective multiorgan donors who did not have cardiovascular pathology in cases in which technical reasons prevented transplantation. Studies were conducted under an institutional review board-approved protocol. This study was approved by the ethical committees of the First Affiliated Hospital of Nanjing Medical University (Protocol Number: 2011-SRFA-075). The investigation conforms to the principles that are outlined in the Declaration of Helsinki regarding the use of human tissues.3 and were fixed with 5% glutaraldehyde in phosphate buffer  at 4°C overnight. After washed in phosphate buffer for four times followed by post-fixation with 1% osmium tetroxide in 0.1 M phosphate buffer for 2 hrs at 4°C, these tissues were dehydrated through graded alcohol at a concentration of 50, 70, 90, 100% for 30 min. for each procedure and embedded in Epon 812. Semi-thin sections were cut at 1.5 μm on a Leica Ultracut R  and stained with toluidine blue and were histologically analysed by using light microscopy. Ultrathin sections were cut at 70 nm and contrasted with uranyl acetate and lead citrate and were examined by using a JEM-1010 electron microscope . The density of TCs was expressed as the mean of the total number of TCs/total area.Heart valves were cut into small pieces of 1 mmFrozen sections were sliced into a thickness of 6 μM, and then were post-fixed with 4% paraformaldehyde dissolved in 0.1 M phosphate buffer (pH = 7.4) for at least 15 min. After washed with phosphate buffer for three times, sections were immersed in 10% goat serum for 1 hr. After that, sections were incubated overnight at 4°C with rat monoclonal anti-CD34  and rabbit monoclonal to Vimentin , both were with the dilution of 1:100 in phosphate buffer and permeabilized by 0.25% Triton X-100 at the same time. On the second day, the sections were incubated to goat anti-rabbit labelled with rhodamine secondary antibodies  and goat anti-rat labelled with FITC  diluted 1:200 in phosphate buffer for 1 hr. The sections were stained with 4′,6-diamidino-2-phenylindole . Similar procedures were used in rabbit monoclonal to PDGF Receptor-beta  and rabbit polyclonal anti-C-Kit .Transmission electron microscopy has been shown to be the most precise way to identify TCs ,6. As sh2 in mitral valve, tricuspid valve and aortic valve.Three different double labelling immunofluorescence methods were used to present evidence for the presence of TCs in human heart valves. These were double labelling for CD34 and c-kit, CD34 and vimentin, and CD34 and PDGFR-β. Figure This study identified TCs as a novel distinct population of interstitial cells in human mitral, tricuspid and aortic valves by their typical ultrastructure and immunophenotype. The cellular body of TC is generally small and oval-shaped . The strin vitro. According to the literature, several relevant and potential roles could be proposed. The existence of telopodes and the 3D network indicates that TCs may function in the intercellular communication and regulation [via ectosomes [This study adds novel evidence for the existence of TCs in human. However, the precise function of TCs in human valves remains to be established. It would be interesting to test the function of TCs gulation ,11, eithctosomes ,45 or byctosomes . MoreoveThus, TCs could represent novel target cells for the treatment of cardiovascular diseases related to the dysfunction of heart valves. The existence of TCs in heart valves provides a novel tool for understanding the essential biological process and even the pathological conditions that have not been determined in heart valve biology.In conclusion, TCs are present in human heart valves. Telocytes may form a 3D network inside the cardiac valves, contributing to mechanical support (force distribution) and flexibility. We speculate that adding TCs to artificial heart valve may lead to better functional properties. It remains to be determined how TCs contributes to the ability of valve to re-establish normal structure and function following injury.