Traditional methods of measuring oral health mainly use clinical dental indices and have been complemented by oral health related quality of life (OHRQoL) measures. Most OHRQoL studies have been on adults and elderly populations. There are no systematic OHRQoL studies of a population-based sample of children. The objective of this study was to assess the prevalence, characteristics and severity of oral impacts in primary school children.Cross-sectional study of all 1126 children aged 11–12 years in a municipal area of Suphanburi province, Thailand. An OHRQoL measure, Child-Oral Impacts on Daily Performances index (Child-OIDP) was used to assess oral impacts. Children were also clinically examined and completed a self-administered questionnaire about demographic information and oral behaviours.89.8% of children had one or more oral impacts. The median impact score was 7.6 and mean score was 8.8. Nearly half (47.0%) of the children with impacts had impacts at very little or little levels of intensity. Most (84.8%) of those with impacts had 1–4 daily performances affected (out of 8 performances). Eating was the most common performance affected (72.9%). The severity of impacts was high for eating and smiling and low for study and social contact performances. The main clinical causes of impacts were sensitive tooth (27.9%), oral ulcers (25.8%), toothache (25.1%) and an exfoliating primary tooth (23.4%).The study reveals that oral health impacts on quality of life in Thai primary school children. Oral impacts were prevalent, but not severe. The impacts mainly related to difficulty eating and smiling. Toothache, oral ulcers and natural processes contributed largely to the incidence of oral impacts. Contemporary concepts of health suggest that dental health should be defined in physical, psychological and social well-being terms in relation to dental status ,2. That A number of socio-dental or OHRQoL measures have been developed and used for assessing oral well-being and to describe oral impacts on people's quality of life . GeneralMost studies using OHRQoL to assess oral impacts of the mouth and teeth have been on adults and elderly populations. Few studies have been done on children possibly because no OHRQoL measures designed for use with children existed until recently. A single measure, dental pain, has been used on children in Malaysia  and in SA cross-sectional survey was carried out in a municipal area of Muang district, Suphanburi province, Thailand. The sample was all 1,126 students aged 11–12 years, in the final year class of all primary schools (grade 6) in the area.Data were collected through: a) an interview for oral impacts using the Child-OIDP , by one The protocol of the study was approved by the Ethical Committee of the Ministry of Public Health of Thailand. Primary education and local health authorities as well as all primary schools in the study areas gave permission. Positive consent forms and letters informing parents were sent to parents.Two comprehensive OHRQoL measures specifically for use with pre-adolescent children have recently been developed; the Child Perceptions Questionnaire (CPQ11-14)  and the The procedure for using the Child-OIDP began with a self-administered questionnaire carried out with all children as a group in their classroom. The questionnaire contains a list of all oral problems that children are likely to perceive and also include an open answer for any unexpected perceived problem. It was developed during a pilot study, as a modification from the one used in the original OIDP. Children were asked to identify oral problems that they perceived in the last three months. This step aimed to focus children's attention to their oral health problems and to lead to the oral impacts assessment later. Their answers here were used only as a guide to investigate oral impacts on daily performances in the next step and were referred to when they were asked about the causes of oral impacts in individual interviews. Thereafter, children were individually interviewed, irrespective of their answers at the first step, to assess oral impacts on daily life in relation to 8 daily performances. The 8 performances were: a) eating, b) speaking, c) cleaning teeth, d) relaxing, including sleeping, e) smiling, laughing and showing teeth without embarrassment, f) maintaining emotional state, g) study, including going to school and doing homework and h) contact with other people. The individual interviews were aided by 16 pictures (negative and positive pictures for each performance). If children reported an impact on any performance, the frequency of the impact and the severity of its effect on their daily life were scored. Children were also asked to identify oral problems that in their opinion, caused the impact. The oral problems were identified from the list complied in the first step of the assessment.The oral impact score of each performance is obtained by multiplying severity and frequency scores, 0, 1, 2 or 3 each, in relation to that performance. Therefore scores can range from 0 to 9 per performance. The overall impacts score is the sum of all 8 performances (ranging from 0 to 72) divided by 72 and multiplied by 100. An alternative method of reporting the severity of oral impacts, from the same data set, is to use the 'intensity' and 'extent' of impacts. The intensity refers to the most severe impacts on any of the 8 performances or the highest performance score. It is classified into 6 levels; none, very little, little, moderate, severe and very severe  completed all stages of the survey. 52.4% were male and 47.6% were female. Their mean age was 11.3 years (sd = 0.6). The highest percentage of their fathers were agricultural workers or labourers (34.5%), 30.5% worked in business/private, 27.5% in governmental sectors, 2.1% did not work and 5.4% of children did not have a male guardian. The highest percentage of mothers worked in business/private (38.6%), 24.5% in agriculture, 21.1% in governmental organisations. 14.9% did not work and 1.0% did not have a female guardian.This population had a low level of dental caries: 43.1% were caries free and the DMFT scores ranged from 0 to 12 with a median score of 1.0 and a mean of 1.5 (±1.8). Almost all children (97.0%) had a Community Periodontal Index (CPI) score of 1 or more; 84.2% had calculus. In terms of oral hygiene status, 5.4% had good, 69.1% had moderate and 25.5% had poor oral hygiene. OHI-S scores ranged from 0.5–5.5 with a median of 2.5 and mean score of 2.5 (±0.9), indicating a moderate level of oral hygiene.The prevalence of oral impacts was high; 89.8% of children had experienced some kind of oral impact on their daily life during the past three months. There was no difference between the prevalence of impacts in girls and boys (Chi-square test). Impacts on Eating were the most prevalent 72.9%). The prevalence of impacts on Emotion (58.1%), Cleaning teeth (48.5%) and Smiling (40.1%) were also relatively high. The remaining prevalences of impacts were lower, namely Study (15.4%), Relaxing (14.7%), Contact with people (12.2%) and Speaking (9.9%) ; 16.2% had 1 PWI, 23.3% had 2, 26.9% had 3 and 18.4% had 4 PWIs. Few children had 5 or more PWIs. About 1 in 5 children had severe or very severe intensity of impacts; 18.7% had severe and 2.6% had very severe intensity of impacts.15.9% had very little, 31.1% had little and 31.7% had moderate intensity of impacts . This is surprising in that this was a low caries population in an area with a free accessible school dental service. Although there is no study using OHRQoL index with a population-based sample of 12 year olds to compare with, findings of previous OHRQoL studies suggest that oral impacts are very common in children of this age. In Brazilian adolescent populations, the prevalence of impacts was 32% ,20 and 69.8%. ThiDespite the fact that oral impacts were prevalent in this Thai child population, they were not severe. For example, half of this population had Child-OIDP score less than 7.6 and half of those with impacts had very little or little intensity of impacts Table . MoreoveThis study found that eating was the most important aspect of OHRQoL of children. Difficulty with eating due to oral problems was the most common impact (72.9%), and led to more severe oral impacts on children's quality of life than impacts on other performances. Oral ulcers and exfoliating primary teeth contributed to eating difficulties in nearly half of those with impacts. The finding that eating was the most common performance affected is similar to all studies using the OIDP in all age groups ,21-24. TDifficulty with smiling was another important aspect of children's OHRQoL. It affected 40% of children. The most prevalent cause was position of teeth. Dissatisfaction with position of teeth, moreover, accounted for oral impacts in 1 in 5 of all children Table . AlthougGum problems were the other important oral conditions affecting children's OHRQoL. More than one fifth of children perceived that bleeding and swollen gums caused oral impacts on their life, particularly in relation to difficulty cleaning, a problem experienced by nearly half of all children Table . ChildreAn interesting finding was that impacts relating to social dimensions, such as study being affected and contact with people, were less common and least severe. Schor suggested that children's social performances rely more on their physical and psychological performances than adults .It is apparent that an important reason for the high prevalence of oral impacts in children is natural processes such as exfoliating primary teeth or spaces due to a non-erupted permanent tooth. They contributed largely to the high incidence of impacts in these pre-adolescent children. On the other hand, these conditions were not reported as important causes of oral impacts in other age groups ,22. The Although children could often not specify precisely which impairments led to impacts, the question of perceived clinical causes should exclude impacts from some conditions which are definitely not related to actual impairments as well as to treatment needs. For example, toothache, ulcers and conditions relating to appearance definitely require different treatment and could be easily differentiated. However, the accuracy of detecting perceived impairment is limited in a population-based study, while it can be improved at the individual level of investigation.The specific age group under investigation, particularly in relation to their stage of development, may have influenced the high prevalence of oral impacts. Developmental changes unavoidably affect HRQoL between childhood and adolescence . MaturitThe prevalence of oral impacts on daily performances in this child population was very high. Oral impacts affected children's quality of life mainly through difficulty eating and smiling. There are various oral conditions that contributed significantly to the incidence of impacts, namely, sensitive teeth, toothache, oral ulcers and exfoliating primary teeth. Although the prevalence of impacts was high, the severity was not; many children had their quality of life affected at low levels. This reveals a need for further longitudinal studies to better understand and interpret OHRQoL measures in children.SG carried out all work including data collection, data analysis and writing the paper. GT supervised the project and assisted writing. AS initiated the idea of, and supervised the project and edited writing.


Recombinant growth hormone preparations have been examined by several methodologies. In this study recombinant human growth hormone ; deamidation of N149, and N152, and oxidation of M140, M125 and M170. Peak area comparison of the modified and parental peptides indicates that these changes were present in ~2% of the recombinant preparation.Electrospray LC-MS analysis revealed one major protein with an average molecular mass of 22126.8 Da and some additional minor components. Electrospray LC-MS/MS evaluation of the enzymatically digested Genotropin70 remains unclear.Modifications of the recombinant human growth hormone may lead to structural or conformational changes, modification of antigenicity and development of antibody formation in treated subjects. Amino acid exchanges may be caused by differences between human and E. coli codon usage and/or unknown copy editing mechanisms. While deamidation and oxidation can be assigned to processing events, the mechanism for possible di-methylation of K The suspension was transferred into Ultrafree-4 centrifugal filter units , for desalting and concentrating proteins. Protein content of the supernatant was quantified by the Bradford protein assay system [Genotropin y system . The staOne dimensional SDS-polyacrylamide gel was performed as described by Laemmli . Samples2 DE was performed essentially as reported . SamplesElectronic images of the gels were recorded using Adobe Photoshop and Microsoft Power Point Softwares.+ ions of angiotensin I, angiotensin II, substance P, bombesin, and adrenocorticotropic hormones (clip 1–17 and clip 18–39). Each spectrum was produced by accumulating data from 200 consecutive laser shots. Those samples which were analysed by PMF from MALDI-TOF were additionally analysed using LIFT-TOF/TOF MS/MS from the same target. A maximum of three precursor ions per sample were chosen for MS/MS analysis. In the TOF1 stage, all ions were accelerated to 8 kV under conditions promoting metastable fragmentation. After selection of jointly migrating parent and fragment ions in a timed ion gate, ions were lifted by 19 kV to high potential energy in the LIFT cell. After further acceleration of the fragment ions in the second ion source, their masses could be simultaneously analysed in the reflector with high sensitivity. PMF and LIFT spectra were interpreted with the Mascot software . Database searches, through Mascot, using combined PMF and MS/MS datasets were performed via BioTools 2.2 software (Bruker). A mass tolerance of 100 ppm and 2 missing cleavage sites for PMF and MS/MS tolerance of 0.5 Da and 1 missing cleavage sites for MS/MS search were allowed and oxidation of methionine residues was considered. The probability score calculated by the software was used as criterion for correct identification.Spots were excised with a spot picker , placed into 96-well microtiter plates and in-gel digestion and sample preparation for MALDI analysis were performed by an automated procedure  ,42. BrieThe algorithm used for determining the probability of a false positive match with a given mass spectrum is described elsewhere .® MiniQuick 0.6 mg  was suspended in the solution provided in the two-chamber cartridge and diluted with 1% formic acid  in water  to 1 pM/μl. 1 μl of this solution was used for the nano-electrospray LC-MS investigation. The HPLC used was an UltiMate™ system  equipped with a PepMap C18 purification column (300 μm × 5 mm) and a 75 μm × 150 mm analytical column of the same material. 0.1% TFA  was used on the Switchos module for the binding of the peptides and a linear gradient of acetonitrile  and 0.1% formic acid in water was used for the elution. The gradient was  0% B for 12 min, 80% B in 30 min, 100 % B in 3 min, 100% B for 10 min, 0% B in 2 min, 0% B for 23 min. The flow rate was 240 nl/min. The LC-system was coupled on-line to a QSTAR Pulsar hybrid mass spectrometer . The nanospray source of Proxeon  was used with the distal coated silica nanospray capillaries of New Objective . The electrospray voltage was set to 1800 V. Spectra were acquired over the mass range of m/z 600–1600. The accumulation time was 1 sec. Protein spectra were deconvoluted by Analyst® . LC-MS/MS analyses were carried out also with the UltiMate™ system interfaced to the QSTAR Pulsar or to an LTQ  linear ion trap mass spectrometer. The gradient was (A = 5% acetonitrile / 0.1% formic acid in water B = 80% acetonitrile / 0.1% formic acid in water) 0% B for 12 min, 60% B in 88 min, 100 % B in 5 min, 100% B for 10 min, 0% B in 5 min, 0% B for 20 min. Peptide spectra were recorded over the mass range of m/z 450–1300, MS/MS spectra were recorded in information dependent data acquisition over the mass range of m/z 50–1600. One peptide spectrum was recorded followed by two MS/MS spectra on the QSTAR Pulsar instrument; the accumulation time was 1 sec for peptide spectra and 2 sec for MS/MS spectra. The collision energy was set automatically according to the mass and charge state of the peptides chosen for fragmentation. One full spectrum was recorded followed by 3 MS/MS spectra on the LTQ instrument, automatic gain control was applied and the collision energy was set to the arbitrary value of 35. Doubly or triply charged ions were selected for product ion spectra. MS/MS spectra were interpreted by Mascot .GenotropinThe author(s) declare that they have no competing interests.Felizardo, Maureen carried out MALDI-TOF-TOF analysis. Raja, Karlin participated in sequence alignments and in the design of the study. All authors read and approved the final manuscript.


Human resources are the most important assets of any health system, and health workforce problems have for decades limited the efficiency and quality of Latin America health systems. World Bank-led reforms aimed at increasing equity, efficiency, quality of care and user satisfaction did not attempt to resolve the human resources problems that had been identified in multiple health sector assessments. However, the two most important reform policies – decentralization and privatization – have had a negative impact on the conditions of employment and prompted opposition from organized professionals and unions. In several countries of the region, the workforce became the most important obstacle to successful reform.This article is based on fieldwork and a review of the literature. It discusses the reasons that led health workers to oppose reform; the institutional and legal constraints to implementing reform as originally designed; the mismatch between the types of personnel needed for reform and the availability of professionals; the deficiencies of the reform implementation process; and the regulatory weaknesses of the region.The discussion presents workforce strategies that the reforms could have included to achieve the intended goals, and the need to take into account the values and political realities of the countries. The authors suggest that autochthonous solutions are more likely to succeed than solutions imported from the outside. Health reforms that aim at increasing efficiency, quality and users' satisfaction need to take into consideration human resource issues, because the health sector is labor-intensive and the performance of health systems depends on qualified and motivated workers -4. At thIn Latin America, the need to improve the performance of the workforce had been pointed out in many health sector assessments conducted in the 1970s and 1980s by the United States Agency for International Development (USAID), the World Bank (WB), other agencies and independent researchers.  were too often guided by favoritism, political dictates, and nepotism;• Health professionals were insufficiently committed to the public system due to the conflict of interests mentioned above, poor personnel management systems and the perception that wages were low;• The medical profession strongly dominated the definition of health sector policies and the regulation of the conditions of practice of all health professions;• Communication between providers and patients was poor, and providers and service users had very different social and cultural backgrounds. In countries with Amerindian-speaking populations, providers did not speak their languages;• The regulation of training institutions and conditions of practice was weak;• The training of health promoters and other auxiliary personnel such as dental assistants, midwives, laboratory technicians, equipment maintenance and repair technicians, and pharmacy clerks was poor or non-existent, thus their performance was poor.According to the literature reviewed, these conditions led to low productivity and efficiency; inadequate equipment; shortages of supplies and drugs; unmotivated and inadequately trained staff; questionable quality of care; and low users' satisfaction.By the mid-1970s, the need to reform the human component of the health services was very urgent, and the urgency increased with the severe economic downturn that countries of the region suffered during in the early 1980s. The size of the Latin America health labor force (about nine million ) impliedThis paper reviews the impact of the health reforms carried out under the leadership of the World Bank. Data come from a review of the literature including the leading Latin American and non-Latin American journals, monographs, documents found in ministries and reform offices, technical reports, papers presented at conferences and fieldwork carried out by the authors between 1980 and the present in Bolivia, Colombia, Costa Rica, Dominican Republic, Ecuador, El Salvador, Honduras, Mexico and Peru.In the early 1980s, with a few exceptions, countries in the region entered a severe economic downturn. The International Monetary Fund and the World Bank required – as a condition for new lending and for refinancing the debts – reduction of public spending. The impact of structural adjustment programs on health services was severe and compromised the ability of governments to maintain the physical infrastructure of the facilities, provide the necessary supplies and equipment, and maintain competitive salaries for the workforce.The World Bank took advantage of this situation to provide loans to the ministries of health and social security funds. Together with the loans, the Bank offered guidelines for the reorganization of the health services according to the ideological economic principles held by the institution.The World Bank had started its activities in the health sector in the early 1970s, and by the early 1990s it was the world's largest health sector lender. In 1999, the total accumulated value of health, nutrition, and population loans worldwide amounted to USD 16.8 billion (in 1996 dollars) . By the The underlying principles of the Bank-led reforms include the belief that the private sector is more efficient than the public sector, and that decentralized administrative units are better-equipped to respond to the needs of the population than centralized governments. The Bank also proposes to limit government financing to a basic package of services, to be determined for each country by cost-effectiveness studies and the country's ability to pay. Services included in the basic package are made available at no cost to the indigent population; governments and the rest of the population subsidize them. In the Bank's health reform model, the role of the state is limited to that of a regulator of the health care market. The United States health system is closer to the World Bank model than the national health services or social security funds of other industrial nations.The World Bank-led reforms did not include strategies to correct the human resources problems presented earlier, even if some of them had been identified by Bank-sponsored studies; the World Bank reformers believed that market forces would resolve them. As we will see, the contrary occurred and privatization and decentralization had some negative consequences for the workforce. The reliance on the market also hid the structural problems that needed to be taken into account at the time reformers designed human resource policies. The design and implementation process which were characterized by secrecy, generated dissatisfaction among health workers.We have organized our analysis of human resources during the reforms under five categories: resistance of the workforce to the implementation of a market-oriented health care model; faulty implementation processes; inadequately trained personnel in managerial positions and in service delivery; institutional and legal dimensions including insufficient and faulty information, lack of financial resources, and civil service statutes; and weak regulatory legislation to ensure the quality of professionals and the performance of the sector.The neoliberal health reforms intended to change the values that had inspired the Latin American systems and the relations between the government and the health workforce. Thus, health would no longer be considered a right; only the insured would be entitled to receive a broad array of services. Health workers would lose the protection they had enjoyed as public servants and become part of a flexible workforce Table .Humans tend to resist change, but opposition to health reform was compounded by conflicts with the value system that inspired it. Most Latin American constitutions recognize health as a right and most governments had adopted Alma Ata's primary health care principles as their strategy to promote Health for All. Health policies were based on the principle of solidarity and fostered cooperation among health workers and also between them and other workers in related sectors such as education and agriculture . It was Human resource changes were needed to implement Alma Ata, but the budget reductions required by the World Bank caused a significant deterioration of working conditions. As their purchasing power worsened, health workers intensified undesirable behaviors to increase their income. These included levying illegal fees, diverting patients from the public sector to their private clinics , using pAs indicated, Latin American physicians have supplemented their income from the public system by means of their private practice. Physicians value the stability and fringe benefits of a public post, but as the public delivery system deteriorated in many countries, private clients became their main source of revenue. By the late 1980s in most Latin American countries , more than half of the health expenditures, including the cost of medicines, occurred in the private sector , and in One of the objectives of the neoliberal reforms was to create a more flexible labor force by decreasing the number of tenured public employees and increasing the number of temporary personnel. This change threatened: the job security of the civil service, a very important dimension in countries with little political stability and where workers are frequently exposed to arbitrary political removals; the providers' income, which now would depend on their ability to compete for contracts and clients; and that which workers expected from their employer . The promoters of health reforms failed to acknowledge that in a politically unstable region, civil service tenure was necessary to maintain an efficient, productive, and loyal labor force. Predictably, these threats triggered the opposition of professional associations and unions, led to strikes, and lowered productivity during the reform process.Health workers of most countries of the region are unionized . The uniLabor unions anticipated that the decentralization and privatization of the sector would have a negative impact on their membership and their bargaining power. Labor unions in Bolivia, Ecuador, El Salvador, Mexico, Nicaragua, Peru and Venezuela expressed their opposition to the reforms on the grounds that they were an excuse for governments to relinquish their constitutional responsibility to ensure access to care and would lead to dramatic changes in work conditions . In someAnalysts of the health reform policies agree with the workers' concerns. Segall  asserts In addition, health professionals strongly opposed the idea of having their professional autonomy limited by professional administrators who would force them to adhere to diagnostic and treatment protocols based on economic principles rather than on technical criteria. The cool reception that Mexican professional associations offered to foreign insurance companies and health maintenance organizations is a good example of this concern .The large majority of countries in Latin America do not have accurate information on the numbers and distribution of the health workforce. There is no centralized entity responsible for gathering information on the health personnel practicing in the country or in the different geographical regions, and even when only public sector workers are examined, the numbers collected at different administrative levels differ . The reforms worsened the possibility of having accurate information. If the system is decentralized and the majority of providers are in the private sector, the sources of information from which human resources data will need to be collated multiplies. The challenge may be even greater if each agency gathers the information differently, and without this basic information it is very difficult to engage in human resources development planning Table .Civil service laws and negotiated agreements with worker unions have limited the freedom of public sector managers to reorganize the workforce; managers had to "administer the rules"  issued eManagers overcame these constraints by hiring additional personnel under flexible contracts. In Brazil there are now about 15 different types of contracts for public sector employees; a significant expansion of the use of temporary workers and contracts without fringe benefits has been reported in Argentina , ColombiTransferring human resources to other politico-administrative levels was more difficult than anticipated. Kolehmainen-Aitken  and HomeDecentralizing human resources is expensive. Prior to decentralization – in countries such as Bolivia, Colombia and Mexico – local or regional governments frequently hired additional personnel under different pay scales and benefit packages than those used by the central government. After decentralization, each decentralized administrative unit had to set pay scales, fringe benefits, performance assessments and reward systems for its workers. Because it was not possible to lower the salaries and reduce the benefits of workers with higher salaries and benefits – generally federal/national workers – it was necessary to bring the salaries and benefits of all the workers to the level of those who had the most generous package. Higher salaries and benefits represent an increase in the fixed costs of the system for an indefinite period.Moreover, the creation and/or strengthening of the decentralized administrative structures are often not accompanied by a decrease in the number of federal employees, further raising the costs of personnel to the system . In MexiDecentralization has been promoted under the assumption that the proximity of decision makers to the communities facilitates providing services more in accordance with the needs of the community. But the decentralized structures' ability to respond to local needs has been constrained by: the civil servants they inherited, who are often inadequate in terms of skill mix and geographical distribution; the conditions of employment imposed by other government sectors and the unions; and the local elite, who do not have in mind the health needs of the communities and who lobby to have relatives and friends hired by local health authorities.In several countries of the region, decentralization has broadened the urban-rural inequities in the distribution of personnel and in the quality of services -54. RichCountries considered the human resource implications of reform only when they faced resistance from the unions or realized they did not have the financial and human resources required to implement the reform . The heaThe human resources units of the health ministries and the regional and local administrations were inadequately prepared for the reform Table . TraditiThe tasks required by the reform overstretched the capacity of these units. Included among these tasks were: the development of new organizational structures, defining new job descriptions, reassigning responsibilities and designing new reporting systems, establishing performance evaluation methods and assisting all decentralized units to carry out similar duties in their jurisdictions.The decentralization of personnel often unveils rivalries and discontent among personnel who feel unfairly treated. All these issues need to be addressed, negotiated and resolved; and most human resources units were ill-prepared to lead the process . For exaThe mismatch between the abilities of the workforce and the needs of the system documented in the 1980s still prevails. The lack of coordination between training institutions and employers is at the root of the problem, along with weaknesses in the regulation of health professions and the dominance of physician's groups on the policy making process.According to Bach , shortagIn most countries, managerial positions were traditionally given to physicians with little or no management training . The neoThe promoters of the reforms recognized the need for good managers but, because they had heavily criticized the public sector and questioned its role, it was difficult to recruit qualified staff into managerial positions and, in turn, it was difficult for those recruited to motivate and retain qualified staff ,57. MostThe Pan American Health Organization  evaluateAfter having spent millions of dollars in training and infrastructure, the region's capacity to manage contracts is still limited , and wheCountries in the region used a top-down approach to define and implement reform. The implementation was often led by a handful of top health executives, newly created reform offices, the political elite and international agencies, which in turn contracted for the technical assistance of international consultants and universities closely aligned with the neoliberal ideology of the World Bank. In general, there was little interest in involving professional associations, unions or even local universities in this process ,56 Tabl.In Costa Rica the labor unions were involved initially in the reform, but their influence was undermined by the World Bank and the Inter-American Development Bank, which adopted a more closed, centralized decision-making style . AccordiSome authors have offered a different interpretation of the exclusion of the workforce and assert that it was the complexity of the human resources issues and the need to involve many players  in their solutions that led reform promoters to ignore the labor force and other stakeholders .Regardless of the underlying motivation, the secrecy that surrounded the process of defining and implementing the reforms produced rumors, confusion and hostility against the reform among civil servants and professional groups ,39,67,68In addition, health reformers did not consider the strategies and resources needed during the transition and early stages of implementation; such as allocating new financial resources and establishing clear communication channels. In addition, as discussed in the previous section, the reforms failed to adequately train managers who could lead the transition to a new management system .The need for regulation increases in health systems where the private sector plays a prominent role. In Latin America it was not until the early 1990s, largely as a consequence of the health reforms, that health policy makers became aware of the urgency to regulate the health system. A regulatory system includes adequate regulations, the institutions to enforce them, and a judicial system that ensures a timely response in the event of conflict. Enforcing regulations is important to guarantee that trained personnel provide safe and adequate services Table .Prior to the reforms, the regulatory systems were poorly developed and, when they existed, they were not tailored to the needs of consumers and enforcement was very limited . For insThe number of medical schools has grown spectacularly in the last two decades and, in most countries, there were no mechanisms to ensure the quality of the training institutions or to test the abilities of the graduates. The number of medical schools in Chile grew by 68% between 1992 and 2000, in Peru the growth was also by 68%, in Argentina by 61% and in Brazil by 21%. The growth occurred mainly in the private sector . Costa RBy its very nature the regulation of the health profession relies heavily on the opinions of professionals and especially on physicians, who in turn place a great value on their autonomy and have had little interest in responding to social and political demands . MedicalThe dominance of physicians has alienated other professions such as therapists, nurses, pharmacists, optometrists and psychologists . For exaThe regulation of the health professions has a long way to go in Latin America, and it is probably impossible to establish sustainable regulatory mechanisms in the absence of political and judicial reforms; for the system to work, it needs to free itself from political interference . Some arBach , Brito eThe implementation of the reforms has been uneven in the Latin American region. Technical, logistical, political and financial problems have surfaced everywhere. While most countries decentralized, a few – such as Colombia and Chile – managed to significantly expand private insurance and, with the exception of Brazil, very few have engaged in large contracts with private sector providers. The most salient feature has been significant changes in hiring modalities. In Brazil there are 15 different types of contract arrangements  and in PAnother important result of the reform is the surge of multiple jobs, particularly in Argentina, Brazil, El Salvador, Panama, Peru, Uruguay, and to a lesser degree Chile ; that haA survey of nurses conducted in Argentina, Brazil, Colombia and Mexico  revealedFurthermore, according to the Tavistock Group  "cooperaSimilarly, one of the basic principles of neoliberal reforms – that the efficiency of the system will increase by using flexible contracts and rewarding productivity – is not supported by the data. The World Bank conducted an evaluation of civil service reforms in 15 countries and concluded: "None of the cases reviewed so far have revealed any empirical evidence that the Civil Service Reform and related Technical Assistance Loans have succeeded in fostering the needed change in work attitudes, ethics and organizational culture that could lead to greater efficiency/productivity in the civil service" [Research has also uncovered problems with using performance-based payment schemes. For example, in health it is often difficult to establish who is responsible for the outcome. Costa Rica implemented a pilot project in Barva de Heredia in which physicians received an incentive based on productivity, but this was not extended to the nurses and the other clinic staff . The proHealth providers can also manipulate the information to maximize their benefits rather than the well-being of the patients; and substandard working conditions rather than workers' action may be responsible for a poor outcome . MexicanEstablishing a valid and reliable merit-pay system is extremely complicated; placing too much emphasis on material rewards may displace more intrinsic motivators such as the pleasure of doing good, or caring for the patient. Bennet and Franco  even sugThere is a belief among neoliberal economists that private sector workers are more productive and less corrupt than public employees. Recent hospital studies confirm that because of fear of termination, absenteeism is less frequent among non-tenured physicians hired through short-term contracts than among civil servants, but short-term contracts have not increased commitment to the institution . CorruptDecentralization can also be seen as a transfer of financial responsibilities from central government to local authorities, which has the potential to affect wages and job stability  and incrIn decentralized health systems, particular attention needs to be paid to establishing good coordination among those responsible for the vertical program at the central level, the decentralized administrative units, and the clinical staff. The absence of good coordination may result in health workers' reporting to two supervisors: the person responsible for the vertical program and the supervisor of the health facility or region, as is the case in Mexico .In sum, in the majority of Latin American countries, the neoliberal reforms have not made the health delivery model more responsive to the needs of the community; have not increased the productivity of health workers; have had a negative impact on working conditions and staff motivation; appear to have further compromised the quality of care; and have had a limited impact on the capacity to regulate the health professions and training institutions.More recent studies suggest that many of the old workforce problems remain unresolved ,70,86,87Health reform provided a perfect opportunity to promote and encourage workforce improvement. Prerequisites for the progress of such processes are political will, effective relationships between the educational and service-providing institutions, and the open collaboration of professional groups. However, the reforms had the opposite effect. The neoliberal orientation challenged the use of conventional regulation strategies because, by encouraging professionals to seek their own interest instead of the interests of society as a whole, it questioned whether society and regulatory bodies could continue to trust and have faith in the criteria expressed by health professionals.Human resources account for the lion's share of health budgets, and poor performance had been identified as the main constraint to efficiency, quality of services and user's satisfaction. The values that guided the neoliberal reform and the privatization and decentralization initiatives worsened the problems affecting the Latin American workforce and added new challenges. The reform implementation process was also responsible for the failures. The promoters of the neoliberal health reforms underestimated the importance of involving professional organizations and unions in the planning and implementation of the reform efforts, raising suspicion and resistance among organized health workers. In addition, the reforms were designed in secret and implemented using a top-down approach. The only significant advance in human resource development in the last 10 years is the increased interest in strengthening the capacity to regulate training institutions and practitioners.Solving the problems that affect human resources is no easy task, and probably the solutions are different for each country; ignoring the problems and hoping that the market will resolve them is a recipe for failure. Managing change is very complex; embarking on reform without having secured the collaboration of the workforce and ensuring the availability of sufficient qualified staff is irresponsible.The exclusion of the organized labor force from reform discussions is indefensible; the collaboration and motivation of the health workers is essential to the reform, and the leaders of the organized groups can assist in informing and aligning the workforce ,89.Policy changes requiring a different skill mix of health personnel require careful planning because there is a significant time lag between deciding that there is a need to train additional professionals and having them available. For example, a 10% rise in the number of students in a medical school produces only a 2% increase in the supply of doctors 10 years later . Most coOne thing that reforms could have done was to support interventions that, independently from the reform, some countries had designed to overcome workforce weaknesses. Some interventions were national programs, others were pilot projects, and there were also experimental projects. The following are examples of these autochthonous interventions.To solve the urban-rural gap and improve equity, most ministries of health in the region created the obligatory rural health service known as pasantía or servicio social that requires physicians to spend one year in a rural health center prior to graduation or immediately after receiving their degree .A few social security institutes have also found ways to reduce health inequities; such is the case of the Mexican and Ecuadorian Institutes of Social Security (IMSS and IESS). IMSS organized COPLAMAR, an extensive primary and secondary care program for poor rural populations, which brought general practitioners and specialists to rural areas. The Ecuadorian program known as Seguro Social Campesino offered primary care services for rural dwellers and, when needed, hospitalization at the Ecuadorian Social Security Institute; this program aimed at reducing the rural-urban gap in a country of which 70% of the population then lived in the rural areas .Mexico created a training program for traditional midwives who worked in dispersed rural populations; the objective of the program was to enhance the quality of their services and reduce maternal and infant mortality . The MinCosta Rica attracted the world's attention with the Open Walls Hospital, a program that required the specialists of a regional hospital to schedule – when needed – weekly visits to dispersed rural populations. The program also intended to convey the message to specialists that they were not different from other health workers and had an obligation to serve poor rural dwellers even when doing so would involve personal inconvenience .To enhance the professional status of primary care practitioners, reduce referrals, and improve quality of care, IMSS created many positions in the specialty of family physician, forcing the medical profession to recognize the status of the new specialty. Colombia's Ministry of Health sent nurses to a graduate health education program taking advantage of a US fellowship program in order to diversify the human resource composition of the Ministry.Some of these projects were successful; this was the case of family physicians and COPLAMAR in Mexico. However, the first 14 Mexican states that decentralized in the 1980s dismantled the program, transferring it to the states' departments of health to create the state health system, and the quality of rural health deteriorated rapidly .Others function poorly, as is the case of the compulsory year of social service in all the countries where it was established. Similarly, the health promoters program in the Dominican Republic suffered from insufficient training, lack of continuous education, absence of efficient supervision, and poor remuneration, comments that can be extended to all health promoter programs of the region. The Seguro Social Campesino has suffered from inadequate financing, and plans to extend it to the entire rural population have been placed on permanent hold.Other programs were discontinued because of indifference and lack of support from policy makers. Thus, the Hospital without Walls ceased after all Ministry of Health hospitals were transferred to the Social Security Institute. Due to budgetary problems, the Colombian Ministry of Health did not employ the health educators on their return from graduate school.The health reforms would have provided a perfect opportunity to support and strengthen many of these and other autochthonous interventions. A proper course of action would have been to evaluate the projects that had been developed locally, identify their strengths and weaknesses, and establish their viability and sustainability. Through trial and error and with appropriate resources and incentives, many of them are likely to be more effective and less costly than foreign programs invented by those who hardly know the realities of developing countries and are inspired by ideological principles and questionable economic theories.There is much more that needs to be done to improve the training and management of human resources for health, and very often the solutions depend on the collaboration of a wide range of stakeholders such as those who produce health workers, those who employ them, those who pay for their services, those who negotiate working conditions and those who define the standards of professional practice. It is no easy task and can be successfully accomplished only if there is strong political will, if there is openness and trust among all stakeholders, and if sufficient resources and time are allocated to this effort. Most countries of the region have the capacity to find appropriate solutions to the problems they are facing. Dussault  argued tAs Segall mentioned , it is iThe author(s) declare that they have no competing interests.Both authors have contributed equally to the design, data collection, data analysis, drafting and completion of this article.


Most lymphomas that involve the tonsil are large B cell lymphomas. Large B-cell lymphoma is a high grade malignancy which progresses rapidly. Tonsillar lymphoma usually presents as either a unilaterally enlarged palatine tonsil or as an ulcerative and fungating lesion over the tonsillar area. Small lymphocytic lymphomas (SLL) of the Waldeyer's ring are uncommon.We report a 41-year-old male who presented with a ten-year history of snoring. Physical examination revealed smooth bilateral symmetrically enlarged tonsils without abnormal surface change or cervical lymphadenopathy. Palatal redundancy and a narrowed oropharyngeal airway were also noted. The respiratory disturbance index (RDI) was 66 per hour, and severe obstruction sleep apnea (OSA) was suspected. No B symptoms, sore throat, odynophagia or dysphagia was found. We performed uvulopalatopharyngoplasty (UPPP) and pathological examination revealed incidental small B-cell lymphocytic lymphoma (SLL).It is uncommon for lymphoma to initially present as OSA. SLL is an indolent malignancy and is not easy to detect in the early stage. We conclude that SLL may be a contributing factor of OSA in the present case. Adenotonsillar enlargement is the main cause of obstructive sleep apnea (OSA) in the pediatric population. However, this prevalent syndrome is more complicated in adults . OSA has2, 91%, minimal SaO2, 62%, and a desaturation index (≥ 4%) of 61.8/h. The arousal index was 64.8/h and the respiratory disturbance index (RDI) was 66.0/h. Believing that the patient was suffering from severe OSA and hyperplastic palatine tonsils, he received UPPP.A 41-year-old man presented with complaints of snoring, excessive daytime sleepiness, and pavor nocturnes for more than 10 years. Systemic diseases were denied. Physical examination revealed bilateral symmetric and enlarged palatine tonsils without abnormal surface change. There were no palpable cervical lymph nodes or B symptoms . White and red blood cell counts, biochemistry and chest radiographs were within normal limits. The results of an overnight polysomnography (PSG) showed mean SaOThe postoperative course was uneventful and sleep apnea improved. PSG performed 4 months after surgery demonstrated that the RDI had reduced to 23.9/h. Pathology indicated small B cell lymphocytic lymphoma Figure ,2 with bAdenotonsillar enlargement is the leading cause of OSA in the pediatric population  though iSmall lymphocytic lymphoma (SLL) is an indolent but relentless malignancy, with a median survival of about 10 years. Because It usually presents as neck lymphadenopathy in the later stages, SLL is not easy to diagnose in the early stage. The effectiveness of chemotherapy for treating SLL is controversial. Most studies have found no benefit in treating patients until they develop symptoms . Lymphomet al., [et al., [Nolan described a case of adenotonsillar enlargement due to chronic lymphatic leukemia which caused severe OSA . His repet al.,  reportedet al., . Abe et [et al.,  describe[et al., .Tonsillar surgery should be performed even on patients highly suspected of having lymphoma to improve OSA -10. NeckNone declared.YT, YC, CL, WC and MT made substantial contributions to the intellectual content of the paper, in the interpretation of results and in drafting the manuscript. All authors read and approved the manuscript


The control of proliferation, differentiation and survival of normal and malignant cells in the tumour microenvironment is under the control of a wide range of different factors, including cell:cell interactions, cytokines, growth factors and hormonal influences. However, the ways in which these factors interact are poorly understood. In order to compare the effects of multiple variables, experimental design becomes complex and difficult to manage. We have therefore evaluated the use of a novel approach to multifactorial experimental design, the Taguchi methods, to approach this problem.The Taguchi methods are widely used by quality engineering scientists to compare the effects of multiple variables, together with their interactions, with a simple and manageable experimental design. In order to evaluate these methods, we have used a simple and robust system to compare a traditional experimental design with the Taguchi Methods. The effect of G-CSF, GM-CSF, IL3 and M-CSF on daunorubicin mediated cytotoxicity in K562 cells was measured using the MTT assay.Both methods demonstrated that the same combination of growth factors at the same concentrations minimised daunorubicin cytotoxicity in this assay.These findings demonstrate that Taguchi methods may be a valuable tool for the investigation of the interactions of multiple variables in the tumour microenvironment. The control of proliferation, differentiation and survival of normal and malignant cells is under the control of a wide range of different factors. These include cell:cell interactions, immune regulatory factors, hormonal influences, and local environmental influences. However, the way in which these factors interact to regulate the dynamics of the malignant cell population are poorly understood. It is important to identify important factors and the way that they interact in order to rationalise treatment and develop new therapeutic options. However, one of the main problems is the difficulty in designing experiments to compare the effects and interactions of multiple variables. For example, a traditional experimental design to compare seven independent variables at three different concentrations each requires a large number of individual experiments (2187 experiments). The logistical and resource implications of this experimental design make these experiments very difficult to carry out. We have investigated the use of an alternative approach to experimental design, the Taguchi Methods , baculovWe have therefore used a simple and reproducible assay, the MTT assay, to evaluate whether the Taguchi methods can be used to investigate the effect of G-CSF, GM-CSF, IL3 and M-CSF on daunorubicin mediated cytotoxicity in K562 cells.Taguchi methods consist of 3 phases: designing the experiment, running and analysing, and confirming and validating the assumptions. After selecting the variables to be studied, Taguchi methods depend on distributing the factors under study in an orthogonal array, which distributes the variables (factors) in a balanced manner. Examining a typical orthogonal array Table , where eA(BC), the subscript L, which is designated by A, represents the number of experiments that would be conducted using this design, B denotes the number of levels or concentrations within each column which denotes how many levels or concentrations could be investigated, while the letter C identifies the number of columns available within the orthogonal array which indicates how many factors or variables could be included in the experiment [8(27) means that 8 experimental runs are needed to investigate 7 different factors, each of which is set at 2 predetermined levels or concentrations  is simply the range, which could be calculated as the difference between the highest and lowest average response of each level. For example the impact of factor A on the experiment outcome is the difference between A1 & A2. ). The effects of all factors are calculated in the same way, then arranged in a response table, and examined for those factors with the strongest effect (i.e. highest difference Δ), in order to separate them from the weak effects. The breaking point between the strong and weak effects is identified as a change in the pattern of the difference between the ranges around the median.Besides determining the effects of the individual factors, the same technique is used to determine the strength of the impact of interactions on the product of the experiment. The calculations are performed as the previous section. In order to determine the interactions between A and B, the average result of each 2 factors combined must be determined. This is achieved through calculating the values of 4 points: A1B1, A1B2, A2B1, and A2B2, where A1B1 is the average result generated due to the interaction between concentration 1 of both factors, A1B2 is the result of the interaction between concentration 1 of factor A and concentration 2 of factor B, A2B1 represents the interaction between factor A at concentration 2 and factor B at concentration 1, while the fourth point A2B2 is the interaction between both factors at concentration 2. These 4 points are then presented graphically to show the strength or weakness of the interaction. Whether the interaction is weak, mild, or strong depends on whether the two response lines are parallel, converging or intersecting, with intersecting lines indicate a strong interaction, and parallel lines indicate no interactions. Once the strong factors and interactions have been identified, an estimate of their combined effect is calculated and the new experiment is designed according to these assumptions. An experiment is then carried out – referred here to as "confirmation run"- to validate the assumptions upon which the new experiment was based. Conducting a confirmation run and the comparison between the actual and the predicted results is necessary. If however, the confirmation results are disappointing, the planning phase must be re-evaluated and the elements that went into the experiment must be reviewed. A possible cause could be the omission of a key factor from the experiment, for example a powerful interaction was not considered. Another common cause is the setting of factor levels too close together for the experiment. In these situations, the factor is found insignificant during the analysis and is not accounted for in the validation. The confirmation run should include the best or preferred settings for mild and weak influences as well as the strong ones. However, the less influential factors are not incorporated into the prediction equation. The reasoning is that the differences in the average results may be due to experimental variation, and to incorporate their effects could result in an overestimate of the predicted results. This could lead to a disappointing confirmation run when actually the results would have validated the experiment analysis if the predicted results had not been artificially high or low.2-95% air. Cells were plated in 96 well microtiter plates (200 μl) at a density of 3 × 104 cells/ml. Cells were co-cultured in the presence of 0.1 μg/ml daunorubicin.K562 cells were cultured in RPMI-1640 medium supplemented by 10% (v/v) foetal bovine serum, 50 μg/ml penicillin and 25 μg/ml streptomycin at 37°C in a humidified atmosphere of 5% COIn all experiments K562 cells were co-incubated in the presence or absence of cytokines concentrations shown in Table 50 μl of MTT  (5 mg/ml) was then added to each well and incubated at 37°C for 4 hours. The resulting deep blue crystals were dissolved in 0.04 N HCl Isopropyl alcohol, and the absorbance measured using a scanning multiwell spectrophotometer at dual wavelength 570–630 nm. All measurements were performed in triplicates.The % survival was calculated asIn classical experimental design the effect of each factor, each concentration and each interaction is tested independently. In order to investigate the full interactions between 4 factors each at 2 concentrations, requires 81 individual experimental conditions to be performed. This study used 49 combinations. The structure of the 49 experiments is shown in Table 8(27) to investigate the effect of 4 cytokines on the survival of K562 leukaemic cells. This array accommodated 4 factors MCSF, IL-3, GMCSF and GCSF. The interaction between MCSF and the other cytokines was inserted into the array. As the 3rd column was used to examine the interaction between MCSF and IL-3, columns 5 and 6 were used to assess the interaction between the same cytokine and GMCSF and GCSF respectively  was observed8 series , four points were computed A1D1, A1D2, A2D1, and A2D2 Table .A1D1 is the average result of the combined effect of MCSF at a concentration 100 U/ml and GCSF at a concentration of 10 ng/ml.The interaction matrix AXD was then represented graphically Fig  which shAn estimate of the predicted response (μ) based on the selected levels was then computed. The calculations were based on the overall average value (T) and the effect that each of the recommended levels of the strong factors and interactions has on the overall average.μ = T+(A2D2 - T)+(B2 - T)+(A2C2 - T)+(A2B2 - T)-(A2 - T)-(A2 - T)-(A2 - T)-(B2 -T)-(C2-T).The reason for subtracting the individual effects of factors A, B, and C from the effects of A2B2 is that A2B2 is comprised of the effects of factor A, factor B and the interaction itself. Unless the effects of the two factors are subtracted these strong effects would be included twice and resulting in an overestimation of the predicted result.The predicted survival derived from the above prediction equation was 43%. A confirmation run that produces a %survival close to 43%would validate the assumptions of this Taguchi method. The actual confirmation run, in fact, resulted in 39.84% survival rate indicating the success of the Taguchi experiment.Further calculations were performed to determine whether the outcome of other combinations could be predicted from the Taguchi experiment and confirmed by analysis. The results are shown in Table The aim of this study was to evaluate the ability of the Taguchi methods to investigate the effects of several factors simultaneously on the death and/or survival of the malignant cells, and to compare this strategy against a traditional full experimental design.A major finding of the study was that the Taguchi methods predicted the combination of factors that results in the lowest survival of the malignant cells. This agreed with the conclusions of the full experimental design but required only eight individual experiments to pinpoint this combination. However, it must be stressed that the Taguchi methods are not intended to be a replacement for traditional experimental design, but if used as a complimentary strategy can make analysis of complex interactions feasible and practical. In this study, for example, eight individual experiments produced a testable combination which required 49 individual experiments to produce in the traditional experimental design. In more complex systems, only Taguchi methods become feasible. For example, to study 13 factors at 3 different combinations would require 1,594,323 individual experiments at a cost for re agents alone of over 27 million pounds. If Taguchi methods are used, this can be reduced to just 27 individual experiments at a cost of under 500 pounds.in vivo, and we propose that the Taguchi methods may be a useful strategy to understand these interactions in vitro, and to help devise and implement new therapeutic strategies.We have described a novel experimental approach to studying the interactions of several factors on the cytotoxicity of malignant cells. We show that the method is effective in the determination of the optimum conditions, even in the presence of multiple interactions. We anticipate that this experimental strategy will have many applications in the investigation of complex interactions. For example we have used this strategy to model the complex testicular microenvironment and the ability to support the survival of acute lymphoblastic leukaemia cells (manuscript in preparation). These sort of interactions are common in the survival of malignant cells None declared.HM designed the Taguchi assay, carried out cell survival assays, and drafted the manuscript. KLY participated in the design and coordination, and produced the final manuscript. APJ conceived the study and participated in its design and coordination. All authors read and approved the final manuscript.


Identification of effective systemic antineoplastic drugs against anaplastic thyroid carcinomas has particularly important implications. In fact, the efficacy of the chemotherapeutic agents presently used in these tumours, is strongly limited by their low therapeutic index.In this study gemcitabine was entrapped within a pegylated liposomal delivery system to improve the drug antitumoral activity, thus exploiting the possibility to reduce doses to be administered in cancer therapy. The cytotoxic effects of free or liposome-entrapped gemcitabine was evaluated against a human thyroid tumour cell line. ARO cells, derived from a thyroid anaplastic carcinoma, were exposed to different concentrations of the drug. Liposomes formulations were made up of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine/cholesterol/1,2-distearoyl-sn-glycero-3-phosphoethanolamine-MPEG (8:3:1 molar ratio). Cell viability was assessed by both trypan bleu dye exclusion assay and fluorimetric analysis of cell DNA content.A cytotoxic effect of free gemcitabine was present only after 72 h incubation . When gemcitabine was encapsulated in liposomes, a significant effect was observed by using lower concentrations of the drug  and earlier exposure time (24 h).in vitro against human thyroid cancer cells, the gemcitabine incorporation within liposomes enhances the drug cytotoxic effect with respect to free gemcitabine, thus suggesting a more effective drug uptake inside the cells. This may allow the use of new formulations with lower dosages (side effect free) for the treatment of anaplastic human thyroid tumours.These findings show that,  At present, the prognosis of anaplastic thyroid carcinomas is very poor ,2. SinceGemcitabine is a new fluorinated nucleoside analogue provided with a potent anti-tumour activity, tested in vitro and in vivo against a variety of solid malignancies -6. In ad1-difluorodeoxycytidine) hydrochloride (HPLC purity >99%) was a kind gift of Eli-Lilly Italia S.p.A.  and was used without further purification. Cholesterol was obtained from Sigma Chemicals Co. . 1,2-dipalmitoyl-sn-glycero-3-phosphocholine monohydrate and N-(carbonyl-methoxypolyethyleneglycol-2000)-1,2-distearoyl-sn-glycero-3-phosphoethanolamine sodium salt (MPEG-2000-DSPE) were purchased from Genzyme products . Double-distilled pyrogen-free water from Sifra S.p.A.  was used. Sterile saline was a product of Frekenius Kabi Potenza S.r.l. . All other materials and solvents were of analytical grade .Gemcitabine . To maximize the entrapment efficiency of a hydrophilic molecule such as gemcitabine, various liposome preparation procedures ,12 were y = 6.958 × 10-3 + 0.3971x, where y is the UV adsorbance and x is the drug concentration) presented a r2 value of 0.9993. The amount of gemcitabine entrapped in liposomes was determined as a difference between the amount of drug added during liposome preparation and the amount of the entrapped drug present in the supernatant. The encapsulation efficiency was expressed as percentage of the total amount of gemcitabine that became entrapped.The loading capacity of liposomes was evaluated after removing the free gemcitabine (centrifugation at 14000 × g for 1 h at 4°C) from the vesicular colloidal suspension coming from the re-suspension of freeze-dried liposomes. The amount of gemcitabine in the supernatant was spectrophotometrically determined at 268.8 nm (Shimadzu UV-1601). UV calibration straight-line  before they reach their target site, one of these is the large volume of distribution after i.v. administration. This situation elicits a low therapeutic index and a high toxicity in healthy tissues . After eIn the last years, gemcitabine has emerged as a very potent anti-tumour drug and it is currently used, alone or in combination, in the treatment of patients with different malignancies, including ovarian, pancreatic, non-small cell lung and other cancers -6. Despi®.In this study, we prepared a liposome formulation with a high entrapment efficiency of gemcitabine and tested its efficacy against a well established human poorly differentiated thyroid carcinoma cell line. Our data demonstrate that, in ARO cells, incorporation of gemcitabine in liposomes enhances the cytotoxic effect of the drug as compared to free gemcitabine, suggesting a more effective uptake of the drug inside the cells. This activity was demonstrated at concentrations even lower than serum levels obtained in clinical trials. In our knowledge, no data are available in the literature concerning the relationship between gemcitabine uptake and its cytotoxicity, since neither liposomes or other carrier system have been tested against tumour cells. Our preliminary experiments (unpublished observations) show that a similar effect (improved uptake and cytotoxicity at lower doses) may be obtained also in colon cancer cells. It is note worthing that the various component used for the preparation of the liposomal delivery device were already approved by regulatory offices to be used in the preparation of liposome-based formulations, i.e. DoxilThe clinical predictive value of the in vitro cell line preclinical cancer model is well established  and the None declared.The pre-publication history for this paper can be accessed here:


Tetanus is a serious but vaccine-preventable disease and fatality rate of the disease is high in the neonates and the elderly. The aim of this study was to detect the tetanus antibody prevalence in the over sixty-year age residents of the nursing homes in Bolu.A voluntary-based study was done in the residents of two nursing homes in Bolu, Turkey. Blood samples were taken from 71 volunteers residing in there nursing homes. Tetanus IgG antibodies were measured by a commercial ELISA kit.Among overall subjects, only 11 (15.7 %) had the protective tetanus antibody titers at the time of the study. Totally, 10 subjects were examined in emergency rooms due to trauma or accidents within the last ten years and, four (40%) of them had protective antibody levels. Of the remaining 61 subjects only 7 (11%) had protective antibody levels (p < 0.05) .Tetanus antibody level is below the protective level in the majority of the over-sixty-year-age subjects residing in the nursing homes. Each over sixty-year age person in our country should be vaccinated. Until this is accomplished, at least, nursing home residents should be vaccinated during registration. Clostridium tetani. Tetanus spores are present in soil and manure and may be introduced into the body through a puncture wound, burn or scratch. Tetanus can never be eradicated because the spores are widely spread in the environment, and found in the intestinal flora of domestic animals, horses, chickens, and humans. Tetanus is not spread from person to person  at a 450/620 nm wavelength. The results were evaluated in the way previously defined by Schoroder et al. . BrieflyStudents't-test was used to compare quantitative variables while chi-square and Fischer's exact tests (two-tailed) were used for qualitative data, p < 0.05 was considered to be significant. Statistical analyses were performed with Epi-info 6.0 .We aimed to include all residents of the nursing homes. In the study period, 78 subjects were staying in the two nursing homes . However, 7 subjects did not want to participate in the study. Therefore, these seven subjects could not be included.A total of 71 voluntary subjects [54 (76%) male and 17 (24%) female] were included in the study. The mean age was 71. Only 11 (15.4%) of the 71 subjects had a protective antibody level against tetanus. 15 subjects were aged between 60–65 (3 of them had protective antibodies), 22 subjects were aged between 66–75 (2 of them had protective antibodies) and 34 of 71 subjects were aged ≥ 76 years (6 of them had protective antibodies). Protective antibody level decreased with every age group  above 60 years old, but there were no statistically significant difference between the five-year age periods above 60 (p > 0.05)  of them had protective antibody levels, only 7 (11%) of 61 subjects who didn't get examined in the emergency room had protective antibody levels (p < 0.05) .Only 10 (14%) of the 71 subjects had been vaccinated in the last ten years. Five of them had been vaccinated as a result of emergency room visits. Antibody level and demographic properties of subjects were presented in the Table According to the records of the Ministry of Health, 2039 tetanus cases were detected in Turkey between 1980–2002, 462 (23%) of which died . In our In our study group, only 11 15.4%) of 71 subjects had protective antibody levels while the rest were under the risk of tetanus. This resultsuggested that lack of protective immunity among nursing home residents was a significant public health concern. In industrialized countries tetanus has become a rare disease and an infrequent cause of death, mainly due to the implementation of comprehensive immunization programs. But, tetanus is still an important problem for the developing countries, due to poor immunization standards and inadequate hygiene . Deaths .4% of 71Individuals over 60 are usually retired people and spending most of their time in garden or land-work. Tetanus spores may be introduced into the body through a puncture wound, scratch during these works. So they face risk of getting tetanus. Also, 28 % of our subjects still were busy with soil and garden work.According to our results, antibody levels in individuals who had been examined in an emergency room within the last ten years due to injury are significantly higher than who didn't have such a history (p < 0.05). This situation suggests that vaccination programs are not well established in countries like Turkey, thus injuries and accidents expose people over 60 to a great risk of getting the tetanus prophylaxis .Once the existence of tetanus is suspected, intensive, and effective management is essential. The patient should receive intensive care aimed at prevention of muscle spasms, prevention of respiratory tract and metabolic complications, and neutralization of circulating toxins . AssuminThe lack of protective immunity against tetanus among the nursing home residents is a significant public health concern. A vaccination program including every individual over 60 should be charted immediately. After the primary series of 3 doses, protection against tetanus should be sustained by scheduling booster doses routinely in every 10 years . Until tThe author(s) declare that they have no competing interests.All authors read and approved the final manuscript. OK designed the study and drafted the manuscript. AT, KA, analysed the data. OK oversaw the microbiological research. OK and FO interpreted the results of the analysis and critically reviewed the final manuscript.The pre-publication history for this paper can be accessed here:


Creating a new species is a bit like climbing a greased flagpole—it's hard to get started and even harder to keep going. Random genetic variations may introduce slight differences between two groups but, without some means to keep them apart, sexual interbreeding will quickly remix the genes and obliterate the differences. Accidents of geography—the rising of mountains or a course-changing river, for instance—can provide physical isolation, which then enables genetic divergence through the accumulation of mutations, either through natural selection or genetic drift.In contrast, speciation without geographic separation relies on the direct action of natural selection to complete the speciation process by strengthening behavioral differences, a process called reinforcement. One of the most powerful means of completing speciation is through the evolution of mate discrimination.Drosophila. They identify two loci that influence the likelihood that a female will choose to mate with a conspecific male, rather than one of a closely related species.A study by Daniel Ortiz-Barrientos and colleagues focuses on the genetic underpinnings of mate discrimination in Drosophila pseudoobscura and D. persimilis exist together along the west coast of the United States (sympatry), but separately elsewhere . When together, they hybridize and produce sterile males. While D. pseudoobscura males will court females of both species, females prefer conspecific males. This female preference is stronger in sympatric females, an enhancement that presumably evolved by the direct action of natural selection to prevent females from wasting their reproductive efforts producing sterile sons. This variation allowed the authors to conduct a series of genetic crosses among flies of the same species but from different locations. Because the daughters of discriminating D. pseudoobscura females were just as discriminating as their mothers, Ortiz-Barrientos and colleagues concluded that female mating discrimination was inherited as a dominant trait. Further crosses showed that genes responsible for female preference were on the X and fourth chromosomes, and high-resolution mapping refined their locations sufficiently to allow the identification of likely candidate genes. While more work remains to be done, the most promising genes in both regions appear to be involved with olfaction. The fact that one of them, CG13982, is known to up-regulate the other, bru-3, strengthens the case that these are indeed promising candidates.D. pseudoobscura based on the combined response to auditory and olfactory cues. The first of these two layers, weak, or “basal” mating discrimination, has previously been associated with a set of traits for acoustic recognition and mapped to chromosomal regions that are inverted between the two species. Such inversions prevent recombination from purging alleles, thereby contributing to hybrid male sterility. As a consequence, when these species interbreed, they inexorably produce sterile males. The second layer, elucidated in the current study, is “reinforced” mating discrimination, which appears to be related to olfactory cues. This additional system of discrimination helps the first layer to fully eliminate the inevitable cost of producing sterile males. Once the nascent species have started up that slippery pole, reinforced discrimination could provide the traction needed to reach its top.According to their findings, the authors propose a novel model of mating discrimination in


Implantation of the embryo and successful pregnancy are dependent on the differentiation of endometrial stromal cells into decidual cells. Female interleukin-11 receptor α (IL-11Rα) deficient mice are infertile due to disrupted decidualization, suggesting a critical role for IL-11 and its target genes in implantation. The molecular targets of IL-11 in the uterus are unknown, but it is likely that IL-11 signaling modifies the expression of other genes important in decidualization. This study aimed to identify genes regulated by IL-11 during decidualization in mouse uterus, and to examine their expression and localization as an indication of functional significance during early pregnancy.IL11Ra+/+) and IL-11Rα deficient (IL11Ra-/-) littermates by oil injection into the uterine lumen, and gene expression analyzed by NIA 15K cDNA microarray analysis at subsequent time points. Quantitative real-time RT-PCR was used as an alternative mRNA quantitation method and the expression and cellular localization of the protein products was examined by immunohistochemistry.Decidualization was artificially induced in pseudopregnant wild type , biglycan and nidogen-1 (entactin). Immunohistochemistry confirmed increased collagen III and biglycan protein expression in IL11Ra-/- uterus at this time. In both IL11Ra-/- and wild type uterus, collagen III and biglycan were primarily localized to the outer connective tissue and smooth muscle cells of the myometrium, with diffuse staining in the cytoplasm of decidualized stromal cells.Among 15,247 DNA probes, 13 showed increased and 4 decreased expression in These data suggest that IL-11 regulates changes in the uterine extracellular matrix that are necessary for decidualization. Implantation of the embryo, formation of the placenta and successful pregnancy are dependent on the proliferation and differentiation of endometrial stromal cells into decidual cells. Decidualization occurs in response to endometrial and embryonic signals and is thought to involve complex interactions between ovarian steroid hormones, the uterine extracellular matrix (ECM), growth factors and cytokines (reviewed in ). When iIL11Ra-/-) mice are unable to support implantation of either IL11Ra-/- or wild type embryos. The failure of decidualization between days 4.5 and 10.5 in IL11Ra-/- females is characterized by greatly reduced vascular permeability at implantation sites, areas of hemorrhage, impaired secondary decidual zone formation, absence of mesometrial decidualization and aberrant infiltration of trophoblast giant cells [IL11Ra-/- mice show some mesometrial decidualization. Females homozygous for a hypomorphic IL-11Rα allele also show reduced decidualization, with decreased cell proliferation, progressive degeneration of the deciduae, infiltration of trophoblast giant cells and absence of placental formation [Female mice with a null mutation in the gene encoding interleukin-11 receptor α (IL-11Rα) are infertile due to disrupted decidualization , suggestnt cells . Althougormation . Neitherormation ,9.Interleukin-11 is a multifunctional cytokine, initially described as a bone marrow stroma-derived hematopoietic growth factor . IL-11 sLocalization and expression of IL-11, IL-11Rα and gp130 in human endometrium across the menstrual cycle suggests a role for this cytokine in decidual transformation in preparation for pregnancy -20. LeveThe molecular targets of IL-11 in the uterus are unknown, but it is likely that IL-11 signaling modifies the expression of other genes important in decidualization. This study aimed to identify genes regulated by IL-11 during decidualization by cDNA microarray, and to examine their expression and localization by immunohistochemistry, as an indication of functional significance during early pregnancy.IL11Ra-/-) had been previously generated by gene targeting, and serially crossed more than 10 generations onto a C57BL/6 background [IL11Ra+/+) and IL-11Rα deficient (IL11Ra-/-) mice, which were identified by Southern blot analysis of genomic DNA obtained from tail biopsies [Mice deficient in IL-11 receptor α  and 80 mg/kg ketamine hydrochloride  in sterile phosphate-buffered saline (PBS). Anesthesia was reversed with 250 μg yohimbine hydrochloride and 400 μg 4-amino pyridine  in sterile PBS.IL11Ra+/+ and IL11Ra-/- littermates at 8–12 weeks of age (n = 4 per genotype per time point) were mated with wild type vasectomized males to induce pseudopregnancy, with the day of plug detection designated day 0. At approximately 1400 h on day 3, decidualization was induced throughout both uterine horns of each animal by injection of 20 μl of sesame oil  into the lumen of each uterine horn via a 26-gauge needle inserted just distal to the utero-tubal junction. Mice were necropsied prior to surgery (0 h) or at 18 h, 24 h or 48 h following artificial decidualization. These time points prior to the onset of secondary decidualization were chosen to ensure similar cellular composition of the IL11Ra+/+ and IL11Ra-/- uteri. Whole uteri were cleaned of fat and weighed. A section of each uterus was either fixed in 10% phosphate-buffered formalin overnight or Carnoy's fixative for 2 h and processed to paraffin wax, or snap frozen in liquid nitrogen for subsequent RNA isolation.Female IL11Ra+/+ (n = 4/time point) and IL11Ra-/- (n = 4/time point) mice were statistically analyzed using GB-Stat 6.5 . Following Bartlett's test for homogeneity of variance, uterine weight was used as the dependent variable and genotype and time as the two independent variables in a two factor analysis of variance. Bonferroni multiple comparison testing was used to compare uterine weight across time in each genotype and between genotypes at each time point. A two-tailed p value of less than 0.05 was considered a significant difference.The wet weights of whole uterus from 260:OD280 = 1.8–2.0). Each artificially decidualized IL11Ra+/+ or IL11Ra-/- uterus was processed individually for microarray hybridization. A reference pool of RNA was prepared from wild type unstimulated uteri (n = 16). The experimental design used indirect comparisons between IL-11Ra+/+ or IL11Ra-/- and the reference pool.Total RNA was extracted from whole uterus by acid guanidinium thiocyanate-phenol-chloroform extraction , incorpoIL11Ra+/+ or IL11Ra-/- cDNA (Cy3) and reference cDNA (Cy5) were competitively hybridized to the microarrays in the presence of 1 mg/ml Cot1 DNA , 10 mg/ml salmon sperm DNA (Gibco-BRL) and 10 mg/ml polyadenylic acid (Sigma). Hybridizations were repeated using the alternate dye combinations to account for differential fluorescent dye incorporation. After washing, slides were scanned using an Axon GenePix 4000B microarray reader  and GenePix Pro 4.0 software (Axon Instruments) to generate pairs of 16-bit tagged image file format (TIFF) files. Following manual quality control for hybridization artefacts, red (Cy5) and green (Cy3) mean foreground and median background fluorescence intensity measurements for each spotted DNA sequence were extracted for export to the statistical programming environment R 1.5.1.Total RNA (10 μg) served as a template for the synthesis of aminoallyl-cDNA, which was then coupled to a fluorescent dye ester (Cy3 or Cy5) as described by the manufacturers of the CyScribe cDNA Post Labelling Kit . Microcon-30 size exclusion columns  were used to initially concentrate RNA samples, and to purify the cDNA probes prior to and following the dye-coupling reaction. Slides were printed with sequence-verified duplicate spots of the NIA 15K cDNA clone set  at the AIL11Ra+/+ (WT) and IL11Ra-/- (KO) uterus at time points following artificial decidualization was determined using the normalization and analysis functions of the statistical language R [R and G) from 4 slides were read into R using the read.genepix and init.data commands. The stat.ma function was used to calculate the log-ratios of expression (M = log2R - log2G) and average log-intensity (A = (log2R + log2G)/2) for each spot, and to normalize the red and green channels relative to one another using print-tip loess normalization [MA plots of each slide were used to determine the effectiveness of this normalization method in adjusting for sources of variation arising from dye bias and print-tip effects.Differential gene expression between nguage R  and the nguage R . For eaclization . Diagnost-statistic and B-statistic for each gene, to give a log odds ratio of difference in mRNA expression between IL11Ra+/+ and IL11Ra-/-. Genes with a log odds score of greater than 3  in both replicates were considered to be significantly up- or down-regulated in IL11Ra-/- uterus compared to wild type.Given the indirect dye swap design  of theseIL11Ra+/+ (n = 2) and IL11Ra-/- (n = 2) uterus at 48 h of decidualization and used for microarray analysis, and an additional two samples per genotype prepared in the same way were used for validation of the microarray data by real-time reverse transcription polymerase chain reaction (RT-PCR). All samples (n = 4/genotype) were further purified through an RNeasy Spin Column , and quantitated by RiboGreen Assay  according to the manufacturer's instructions, prior to triplicate reverse transcription reactions.Total RNA samples extracted from Total RNA (1 μg) was reverse transcribed at 46 C for 1.5 h in 20 μl reaction mixture using 100 ng random hexanucleotide primers and 6 IU AMV reverse transcriptase  in the presence of cDNA synthesis buffer (Roche), 1 mM dNTPs (Roche), 10 mM dithiothreitol (Roche) and 10 IU ribonuclease inhibitor . The resulting cDNA mixtures were heated at 95 C for 5 min before storage at -20 C in small volumes to avoid freeze-thawing. Negative controls were performed by omission of reverse transcriptase.IL11Ra+/+ and IL11Ra-/- uterus at 48 h of decidualization, PCR was carried out using a Roche LightCycler. Prior to LightCycler analysis, standard cDNA for each gene of interest was generated using a PCR Express block cycler . A 1 μl aliquot of RT product was amplified in a total volume of 40 μl using 4 μl of 10 × PCR Reaction Buffer , 62.5 μM dNTPs , 10 pmol sense and antisense primers  was added to sterile capillaries to a total volume of 20 μl containing SYBR Green I, dNTPs, Taq DNA polymerase and reaction buffer , supplemented with 5 pmol of specific sense and antisense primers Table  and an oIL11Ra+/+ and IL11Ra-/- uterus. Following normalization, levels of RNA for COL3A1, BGN, SPARC and NID1 in IL11Ra-/- compared to wild type were statistically analysed using the paired t-test function of GraphPad Prism 3.0 . A two-tailed p value of less than 0.05 was considered a significant difference.Triplicate RT reactions for each sample, the standard curve and a no RT negative control were analyzed in the same run, and each run was repeated once. A mean value for the initial target concentration of each sample was calculated using the fit points function of the LightCycler software. The mean concentration of 18S rRNA for each sample was used to control for RNA input, as it is considered a stable housekeeping gene and was not altered in expression between IL11Ra+/+ and IL11Ra-/- uterus collected at 48 h of decidualization, using specific antibodies . The morphology and cellular localization of artificial decidualization in IL11Ra+/+ and IL11Ra-/- uterus was also determined by immunostaining for the decidual marker desmin [To confirm differential expression of collagen III, biglycan, SPARC and nidogen-1 at the protein level, immunohistochemistry was carried out on transverse sections of r desmin . Primaryr desmin , gift frFive micron sections of Carnoy's-fixed  or formalin-fixed  uterus were mounted on poly-L-lysine-coated glass slides, deparaffinized and rehydrated through a series of graded ethanols. In an humidified chamber at 25 C, sections were incubated for 10 min in 3% hydrogen peroxide to block endogenous peroxidase activity, then for 30 min (SPARC and desmin) or 1 h  in 10% serum of the species in which the secondary antibody was raised  and 2% mouse serum in one of the above buffers. Sections were then incubated with primary antibody or negative at 4 C overnight  or 30 min at 25 C (desmin), then washed in TBS/Tween prior to secondary antibody incubation for 30 min (SPARC and desmin) or 1 h  at 25 C. Sections were again washed in TBS/Tween, then the secondary antibody detected using the Vectastain ABC Elite/HRP Kit  or the StreptABComplex/HRP Kit  according to the manufacturer's instructions. Protein localization was visualized using the Liquid DAB-Plus Substrate Chromogen System (DAKO), with Harris hematoxylin (Sigma) counterstain.p < 0.01) at the final time point  in uterine weight at 48 h of artificial decidualization between IL11Ra+/+ (96.9 +/- 9.8 mg) and IL11Ra-/- (30.4 +/- 7.7 mg).Following the injection of oil into the wild type pseudopregnant uterus, a progressive increase in uterine weight was observed from 0 through to 48 h, reaching statistical significance p < 0.0 at the fIL11Ra+/+ and IL11Ra-/- uterus artificially decidualized for 0, 18, 24 or 48 h (n = 2/genotype/time point) was used as a template for the hybridization of NIA 15K cDNA microarrays. Figure p = 0.05). At 0 h, prior to application of the decidualizing stimulus on day 3 of pseudopregnancy, there were no reproducible differentially expressed genes between IL11Ra+/+ and IL11Ra-/- , COL3A1  and IL11Ra-/- (1.66 +/- 0.09 fg/μl) uterus approached statistical significance at p = 0.0708.To confirm the altered mRNA expression of the ECM genes COL3A1, SPARC, BGN and NID1 at 48 h of decidualization, quantitative real-time RT-PCR was carried out using the same RNA samples used in the microarray analysis, plus two additional RNA samples of each genotype, collected in the same way. At a significance level of 3A1 Fig. , SPARC . GrowthIL11Ra-/- compared to wild type uterus, this effect was not statistically significant when real-time RT-PCR was used as an alternative quantitation method. Many factors may contribute to discrepancies between cDNA microarray and real-time RT-PCR data. There are major differences in the approach to mRNA quantitation used by the two techniques. Using cDNA microarray, the mean fluorescence intensity ratio for each gene in IL11Ra-/- or IL11Ra+/+ uterus was calculated relative to a reference pool, and the ratio of IL11Ra-/- to IL11Ra+/+ determined by the use of computational algorithms. When quantitating the same mRNA species by real-time RT-PCR, a standard curve of known concentration was used to infer the absolute abundances of mRNA in the IL11Ra-/- and IL11Ra+/+ samples, which were then normalized for RNA input.While the microarray data showed consistent, reproducible upregulation of COL3A1, BGN, SPARC and NID1 in Real-time RT-PCR was chosen for cDNA microarray validation in this study because it has higher sensitivity and lower RNA requirements than Northern blot, but the lack of agreement between the two methods is not unusual. It is well recognized that fold change values for a given gene may vary widely, even between two different microarray techniques -43. In uThere are a number of compelling arguments both for and against conducting corroborative studies for microarray data, and there is good evidence that the data is highly reliable when the experimental design and statistical analysis is sound . In asseIL11Ra-/- uterus is supported by existing data in the literature. Collagen III is a fibrillar collagen with known roles in differentiation and migration [For each of the genes examined, upregulation during defective decidualization in igration , and togigration . Consistigration , and secigration . The redigration  may faciigration . This deigration . In the igration . This isIL11Ra-/- mice could contribute to impaired decidualization.During the human menstrual cycle, collagen III immunostaining is higher in the proliferative compared to the secretory phase , indicatIL11Ra-/- uterus at 48 h of decidualization, it appears that successful decidualization involves downregulation of COL3A1 transcription.Using microarray analysis, the mRNA encoding procollagen III α1 (COL3A1) has been previously shown to decrease in abundance in the mouse uterus at estrus , and betIL11Ra-/- mouse uterus was associated with the upregulation of biglycan mRNA, the activity of this proteoglycan in the ECM may inhibit the decidual response.Biglycan is a small leucine-rich proteoglycan, which binds to type I  and typeDecidual cells are known to express nidogen-1 as part of the pericellular basement-membrane laid down during decidualization . The maiSPARC (osteonectin/BM-40) is described as a matricellular glycoprotein, in that it binds to both cells and ECM to regulate cell-matrix interactions . Like otUsing mRNA and protein expression studies alone, it is not possible to determine whether IL-11 affects ECM molecule expression directly or indirectly. The presence of STAT binding sites in the promoters for COL3A1, BGN, NID1 and SPARC would confirm that a direct interaction is possible, but would not establish that the interaction is occurring in the uterus during decidualization. Given that IL-11 is a secreted cytokine with autocrine and paracrine activity, a direct effect on ECM molecule transcription would be dependent on coincident expression patterns of the ECM molecules, IL-11 and its receptors. Maximal expression of IL-11 and IL-11Rα mRNA has been reported in the predecidual and decidual cells of the mouse implantation site by in situ hybridization . ExpressIL11Ra-/- uterus compared to wild type. Previous in vitro studies have indicated that IL-11 inhibits MMP-1 and -3 protein in human synovium [Indirect effects of IL-11 on ECM composition could be mediated by matrix metalloproteinases (MMPs) and/or their inhibitors. Despite their presence on the NIA 15K microarray, differential expression was not observed for MMP-2, MMP-9, TIMP-2, or TIMP-3 in the synovium , and enhsynovium . IL-11 dsynovium , or indusynovium , but tissynovium . While nThis investigation of the downstream targets of IL-11 during mouse decidualization has uncovered previously unknown interactions between IL-11 and uterine ECM composition. Dysregulation of collagen III, biglycan, nidogen-1 and SPARC in the absence of IL-11 signaling at the time of decidualization may indicate essential functions for these molecules during the implantation process in mice. Functional studies using mouse and human endometrium may further clarify the mechanisms of IL-11 action on the ECM during this critical time in embryo implantation. By elucidating the role of IL-11 regulated genes in decidualization, future work may identify potential new targets for the manipulation of human fertility.IL11Ra+/- breeding pairs. LAS conceived of the study, participated in its design and coordination and helped to draft the manuscript. All authors read and approved the final manuscript.CAW carried out all the experimental work and statistical analysis and drafted the manuscript. LR provided the heterozygote