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Transplantation of pancreatic islets is a promising approach to controlling glucose levels in type 1 diabetes mellitus (T1DM), but islet survival is still limited. To overcome this, islet co-culture with mesenchymal stromal cells (MSCs) together with safe immunosuppressive agents like squalene-gusperimus nanoparticles (Sq-GusNPs) may be applied. This could support islet survival and engraftment. Here, we studied how Sq-GusNPs and adipose-derived stem cells (ASCs) influence islets response under pro-inflammatory conditions. Through qRT-PCR, we studied the expression of specific genes at 24 hours in human and rat islets and ASCs in co-culture under indirect contact with or without treatment with Sq-GusNPs. We characterized how the response of islets and ASCs starts at molecular level before impaired viability or function is observed and how this response differs between species. Human islets and ASCs responses showed to be principally influenced by NF-kappa B activation, whereas rat islet and ASCs responses showed to be principally mediated by nitrosative stress. Rat islets showed tolerance to inflammatory conditions due to IL-1Ra secretion which was also observed in rat ASCs. Human islets induced the expression of cytokines and chemokines with pro-angiogenic, tissue repair, and anti-apoptotic properties in human ASCs under basal conditions. This expression was not inhibited by Sq-GusNPs. Our results showed a clear difference in the response elicited by human and rat islets and ASCs in front of an inflammatory stimulus and Sq-GusNPs. Our data support the use of ASCs and Sq-GusNP to facilitate engraftment of islets for T1DM treatment.

Gusperimus is an anti-inflammatory drug that has shown to be effective in managing autoimmunity and preventing graft rejection. This is unstable and easily broken down into cytotoxic components. We encapsulated gusperimus binding it covalently to squalene obtaining squalene-gusperimus nanoparticles (Sq-GusNPs). These nanoparticles enhanced the immunosuppressive effect of gusperimus in both mouse macrophages and T cells. The half-maximal inhibitory concentration in macrophages was 9-fold lower for Sq-GusNPs compared with the free drug. The anti-inflammatory effect of the Sq-GusNPs was maintained over time without cytotoxicity. By studying nanoparticles uptake by cells with flow cytometry, we demonstrated that Sq-GusNPs are endocytosed by macrophages after binding to low-density lipoprotein receptors (LDLR). In presence of cathepsin B or D release of gusperimus is increased demonstrating the participation of proteases in the release process. Our approach may allow the application of Sq-GusNPs for effective management of inflammatory disorders including autoimmunity and graft rejection.

The Andean-Amazon foothills region, shaped by Andean moist forests and Amazon forests in southwestern Colombia, Napo province in Ecuador, and Ucayali Province and Napo Basin in Peru, provides local and global ecosystem services as food, water, world climate regulation, water purification, and carbon absorption. However, it faces major problems of land-use change that are exacerbated by climate change that affects these ecosystem services. For instance, conventional agriculture contribute to deforestation, soil degradation, and biodiversity loss, which might be further aggravated by climate change–induced droughts, thus reducing staple crop production and, consequently, food security. Cassava (Manihot esculenta Crantz), maize (Zea mays L.), and plantain (Musa paradisiaca L.) are major staple crops in the region. They play a key role for food security and local farmers’ income but are highly exposed to climate risks. This article aims to quantify the level of exposure to climate change (measured as climatic suitability) of these crops in the Andean-Amazon foothills by using the EcoCrop model by the 2030s, 2050s, and 2080s under Representative Concentration Pathway 2.6, 4.5, and 8.5 scenarios. EcoCrop results showed that, whereas cassava will not lose climatic suitability, maize will lose more than half of its current suitable area, and plantain will gain and lose area, which would affect local food security. Globally, these results are important in highlighting adaptive and cost-effective strategies in agriculture and suggest that agricultural crop diversification may improve resilience by promoting the use of local crops varieties.

Grafting of microencapsulated pancreatic islets has been proposed as an alternative to exogenous insulin for the treatment of type 1 diabetes mellitus. Microencapsulated islets are protected from direct contact with immune cells and larger immune-active molecules such as immunoglobulins. Unfortunately, many islet cells in the microcapsules are lost in the immediate period after transplantation due to an early host immune response limiting long-term function of the graft. Gusperimus has shown to reduce the inflammatory responses to grafted encapsulated islets, but it cannot be appropriately used because it is easily hydrolyzed leading to loss of activity. To temporarily modulate the inflammatory response directly after implantation and stabilize gusperimus, squalene-gusperimus nanoparticles (Sq-GusNPs) are developed and incorporated in human islets-containing alginate-based microcapsules. A prolonged and continuous release of gusperimus is achieved. This offers an anti-inflammatory microenvironment in the vicinity of the microcapsules and a reduction of cytokine secretion by lipopolysaccharides-activated human macrophages. Release of gusperimus from Sq-GusNPs does not affect the in vitro viability or function of human pancreatic islets. The data illustrate that incorporation of Sq-GusNPs in alginate microcapsules offers an opportunity to temporarily modulate the immediate immune response after the grafting procedure of encapsulated islets cells and reduce loss of islet cells.

Plants are essential sources of food, medicine, shelter, fuel, feed, and forage, and provide a wide range of additional ecosystem and cultural services to humanity. In recognition of the tremendous value of useful plants and of the increasing threats to their persistence, international agreements including the Convention on Biological Diversity, the Sustainable Development Goals, and the International Treaty on Plant Genetic Resources for Food and Agriculture have created ambitious conservation targets which must be measured through quantitative indicators so as to facilitate the development and implementation of strategies aimed at safeguarding their genetic diversity. Gaps in the current list of functioning indicators for these targets suggest that the development of effective measurements of the state of conservation of the genetic diversity within useful plants is a major challenge. Here we present a gap analysis indicator methodology that provides a pragmatic estimate of the comprehensiveness of conservation of the genetic diversity within useful wild plants, both ex situ and in situ. The methodology compares the geographic and ecological variation evident from analyses of the ‘site of collection’ of samples of plant taxa that are safeguarded in genebanks and other living plant repositories, as well as the variation evident in the proportion of species’ ranges inhabiting protected areas, against the full range of geographic and ecological variation in their native distributions. The methodology enables a prioritization of species for immediate conservation action, and, when measured periodically, can quantify progress toward comprehensive conservation of these plants at global, regional, and national scales, including determining when that goal has been reached. Assessing almost 7000 taxa with the “Comprehensiveness of conservation of useful wild plants” indicator, we find that they are currently highly under-conserved, with less than three out of every 100 taxa assessed as sufficiently conserved or of low priority for further conservation action (overall global indicator = 2.78). Indicator results at the national and regional scales as well as by species use type varied, although virtually all countries, regions, and use categories were found to require further conservation action, particularly with regard to ex situ conservation. The results as well as input data and method code are available for indicator reporting and for conservation prioritization setting.

Zygomatic intraosseous vascular anomalies are uncommon. Most cases have been reported without a precise diagnosis. The prevalence is 3:1 women during the fourth decade of life. The study of these lesions includes clinical findings, radiological studies, such as computed tomography and magnetic resonance imaging scans, and a postoperative histological study of the specimen. Biopsy is not recommended because of the risk of bleeding. Complete surgical resection remains the gold standard elective treatment.The case of a patient with an intraosseous venous malformation of the zygomatic bone is presented, detailing the clinical, histological, and radiological characteristics and the surgical treatment with a primary reconstruction through Computer-Aided Design/Computer-Aided Manufacturing customized titanium implant guided by virtual surgical planning with a reduction of surgical time and morbidity in the donor area with optimal functional and aesthetic results.

BACKGROUND: CIRPLAST is a nonprofit volunteer plastic surgery program that has provided free surgery for patients with cleft lip and palate deformities in different parts of Peru since 1995. In 2015, the author reported 6,108 patients that had been successfully operated on by the CIRPLAST team over a 20-year period. A technique, developed by the author, for the straight-line vertical cleft lip closure without skin flaps of the unilateral cleft lip, was mentioned in that publication but it was not described. 1 The purpose of this article is to present the technique, which has been successfully employed in all the CIRPLAST cleft missions in Peru, for the past 25 years. METHODS: The straight-line vertical cleft closure does not rely on measurements or skin flaps, and it can be used to close any degree of unilateral cleft lip cleft. The procedure is simple and dependable. After incising the cleft borders on both sides of the cleft, the orbicularis oris muscle is liberated from the surrounding tissues, segmented, and then moved down toward the free border of the lip, so that the cupid's bows can be placed in its normal horizontal position, together with the philtrum on the medial lip, providing normal fullness and pouting of the lower part of the upper lip. Lip length results from the orbicularis oris muscle repair and not from skin flaps. The associated nasal deformity is addressed at the same time as the lip repair, by freeing on the cleft side, the lower lateral cartilage (alar cartilage) from the external nasal skin through a rim incision, and then elevating the cartilage together with its vestibular skin, to place it in its normal position at the tip of the nose, and fixing it there with sutures. RESULTS: The anatomic, functional, and esthetic results of the lip closure together with the correction of the associated nasal deformity have been satisfactory, when comparing the repaired cleft side with the normal side, for symmetry. CONCLUSIONS: The straight-line vertical cleft lip closure, based on the orbicularis oris muscle repair, can be used to close any degree of lip clefting, including very wide clefts, without skin flaps. The associated cleft nasal deformity is corrected before the lip closure. The procedure has been used in all the CIRPLAST cleft missions in Peru for the past 25 years, and the outcomes of the repair over time have been satisfactory and stable.

Immunosuppressive drugs are widely used for the treatment of autoimmune diseases and to prevent rejection in organ transplantation. Gusperimus is a relatively safe immunosuppressive drug with low cytotoxicity and reversible side effects. It is highly hydrophilic and unstable. Therefore, it requires administration in high doses which increases its side effects. To overcome this, here we encapsulated gusperimus as squalene-gusperimus nanoparticles (Sq-GusNPs). These nanoparticles (NPs) were obtained from nanoassembly of the squalene gusperimus (Sq-Gus) bioconjugate in water, which was synthesized starting from squalene. The size, charge, and dispersity of the Sq-GusNPs were optimized using the response surface methodology (RSM). The colloidal stability of the Sq-GusNPs was tested using an experimental block design at different storage temperatures after preparing them at different pH conditions. Sq-GusNPs showed to be colloidally stable, non-cytotoxic, readily taken up by cells, and with an anti-inflammatory effect sustained over time. We demonstrate that gusperimus was stabilized through its conjugation with squalene and subsequent formation of NPs allowing its controlled release. Overall, the Sq-GusNPs have the potential to be used as an alternative in approaches for the treatment of different pathologies where a controlled release of gusperimus could be required.

Encapsulation of pancreatic islets in alginate-microcapsules is used to reduce or avoid the application of life-long immunosuppression in preventing rejection. Long-term graft function, however, is limited due to varying degrees of host tissue responses against the capsules. Major graft-longevity limiting responses include inflammatory responses provoked by biomaterials and islet-derived danger-associated molecular patterns (DAMPs). This paper reports on a novel strategy for engineering alginate microcapsules presenting immunomodulatory polymer pectin with varying degrees of methyl-esterification (DM) to reduce these host tissue responses. DM18-pectin/alginate microcapsules show a significant decrease of DAMP-induced Toll-Like Receptor-2 mediated immune activation in vitro, and reduce peri-capsular fibrosis in vivo in mice compared to higher DM-pectin/alginate microcapsules and conventional alginate microcapsules. By testing efficacy of DM18-pectin/alginate microcapsules in vivo, we demonstrate that low-DM pectin support long-term survival of xenotransplanted rat islets in diabetic mice. This study provides a novel strategy to attenuate host responses by creating immunomodulatory capsule surfaces that attenuate activation of specific pro-inflammatory immune receptors locally at the transplantation site.

Climate change will cause geographic range shifts for pollinators and major crops, with global implications for food security and rural livelihoods. However, little is known about the potential for coupled impacts of climate change on pollinators and crops. Coffee production exemplifies this issue, because large losses in areas suitable for coffee production have been projected due to climate change and because coffee production is dependent on bee pollination. We modeled the potential distributions of coffee and coffee pollinators under current and future climates in Latin America to understand whether future coffee-suitable areas will also be suitable for pollinators. Our results suggest that coffee-suitable areas will be reduced 73-88% by 2050 across warming scenarios, a decline 46-76% greater than estimated by global assessments. Mean bee richness will decline 8-18% within future coffee-suitable areas, but all are predicted to contain at least 5 bee species, and 46-59% of future coffee-suitable areas will contain 10 or more species. In our models, coffee suitability and bee richness each increase (i.e., positive coupling) in 10-22% of future coffee-suitable areas. Diminished coffee suitability and bee richness (i.e., negative coupling), however, occur in 34-51% of other areas. Finally, in 31-33% of the future coffee distribution areas, bee richness decreases and coffee suitability increases. Assessing coupled effects of climate change on crop suitability and pollination can help target appropriate management practices, including forest conservation, shade adjustment, crop rotation, or status quo, in different regions.

Device level reliability is introduced into simulations of an optimized all-NMOSFET, cross-coupled, radiofrequency Voltage Controlled Oscillator to trace the sensitive devices and parameters that reduce circuit performance over time. Parametric DC time dependent degradation is considered in all the devices of the circuit along with correlated flicker and thermal noise increase under hot carrier injection stress. Results show that, rather than the changes introduced by stress on DC parameters, the proposed topology is very sensitive to device level noise increase due to stress-induced traps, particularly at early stages of circuit life. This highlights the requirement of a unified simulation framework that takes into account not only DC/AC small signal degradation models but also device noise degradation in noise sensitive circuits.

Projections of climate change are available at coarse scales (70–400 km). But agricultural and species models typically require finer scale climate data to model climate change impacts. Here, we present a global database of future climates developed by applying the delta method –a method for climate model bias correction. We performed a technical evaluation of the bias-correction method using a ‘perfect sibling’ framework and show that it reduces climate model bias by 50–70%. The data include monthly maximum and minimum temperatures and monthly total precipitation, and a set of bioclimatic indices, and can be used for assessing impacts of climate change on agriculture and biodiversity. The data are publicly available in the World Data Center for Climate (WDCC; cera-www.dkrz.de), as well as in the CCAFS-Climate data portal (http://ccafs-climate.org). The database has been used up to date in more than 350 studies of ecosystem and agricultural impact assessment., Measurement(s)climate change • precipitation process • precipitation amount • consecutive dry months index per time period • temperature of airTechnology Type(s)computational modeling techniqueFactor Type(s)spatial regionSample Characteristic - Environmentclimate systemSample Characteristic - LocationEarth (planet) Machine-accessible metadata file describing the reported data: 10.6084/m9.figshare.11353664

To the best of our knowledge, the empirical corpus of work on the origins and persistence of democracy has not evaluated the impact of governments' fiscal independence from peoples' taxes on the quality of democracy. Our proxy for fiscal independence is the component 1C titled "government enterprises and investments" taken from the Economic Freedom of the World index. Component 1C measures in a given country the fraction of total investment that is supplied by the government and state-owned enterprises. A higher fraction corresponds to greater fiscal independence. The econometric analysis employing a variety of IV estimators, which includes Lewbel (2012) generated instruments, reveals a conditional negative, robust, and statistically significant effect of fiscal independence on the degree of democracy. The study also unveils a statistically and economically significant impact of fiscal independence on variables inherent to democracy, such as judicial independence and freedom of the press, as well as on human capital, a proximate determinant of democracy. These findings account for the observed heterogeneity in the potentially confounding effects of democratic rule determinants commonly used in the democratization literature. In particular, the study allows for years of education, ethno-linguistic fractionalization, the individualism-collectivism cultural trait, legal origin, risk of expropriation protection, income inequality, geographical characteristics such as latitude and availability of oil reserves, and fixed effects to control for unobserved heterogeneity bias induced by time-invariant variables at the continent level. The evidence uncovered suggests a novel mechanism under-girding the onset, and more crucially, the persistence of democratic rule. The unveiled finding on fiscal dependency is congruous with landmark historical events such as the English Glorious Revolution as well as the tardier emergence of democracy in continental Europe. Similarly, our findings cast light on the modern heterogeneity of democratic outcomes. Specifically, results inform the flourishing Western democracies experience, and the absence or low levels of democracy among countries where governments are owners of revenue-generating companies or recipients of substantial tax revenues paid by private or state-owned corporations. Conspicuous examples of nations, with governments mostly fiscally independent, are Saudi Arabia and Venezuela.

This research uses predicted genetic diversity unadjusted for the ancestral composition of current populations, as a plausible source of exogenous variations for indicators of economic institutions. While genetic diversity has a robust, concave and significant effect on economic institutions, reduced-form regressions and numerous falsification tests ostensibly suggest that genetic diversity affects development only via indices of multidimensional measures of economic institutions. Second-stage results indicate that allowing for cognitive skills, latitude and ethno-diversity, economic institutions exert a positive and strongly statistically significant effect on development. These findings are robust to the inclusion of deep and proximate growth determinants, different measures of geography, institutions, and horse races between cognitive skills and economic freedom, as well as to the use of different estimators. Human capital, gauged by cognitive skills, in most specifications is not significant in the second stage; however, it is positive and a strong significant predictor of economic institutions in the first stage. The empirical evidence unveiled in this study lends credence to the primacy of economic institutions hypothesis to ignite long-term growth and highlights the crucial role of human capital in enhancing economic institutional quality.

Coffee is grown in more than 60 tropical countries on over 11 million ha by an estimated 25 million farmers, most of whom are smallholders. Several regional studies demonstrate the climate sensitivity of coffee (Coffea arabica) and the likely impact of climate change on coffee suitability, yield, increased pest and disease pressure and farmers’ livelihoods. The objectives of this paper are (i) to quantify the impact of progressive climate change to grow coffee and to produce high quality coffee in Nicaragua and (ii) to develop an adaptation framework across time and space to guide adaptation planning. We used coffee location and cup quality data from Nicaragua in combination with the Maxent and CaNaSTA crop suitability models, the WorldClim historical data and the CMIP3 global circulation models to predict the likely impact of climate change on coffee suitability and quality. We distinguished four different impact scenarios: Very high (coffee disappears), high (large negative changes), medium (little negative changes) and increase (positive changes) in climate suitability. During the Nicaraguan coffee roundtable, most promising adaptation strategies were identified, which we then used to develop a two-dimensional adaptation framework for coffee in time and space. Our analysis indicates that incremental adaptation may occur over short-term horizons at lower altitudes, whereas the same areas may undergo transformative adaptation in the longer term. At higher elevations incremental adaptation may be needed in the long term. The same principle and framework is applicable across coffee growing regions around the world.

This paper offers an overview of how climate change is already affecting farmers across eastern and southern Africa, and how it will continue to affect them in the future. The rising temperatures and increased rainfall variability associated with climate change are undermining the livelihoods and food security of Africa’s farmers, most of whom work at a subsistence level and also face problems of poverty, inadequate infrastructure and poor governance. To address these problems, governments and development organizations have promoted climate-smart agriculture (CSA). These projects, however, have been constrained by inadequate data and predictions regarding future climate change. In particular, farmers in Africa need better projections of the climate hazards for specific regions. Historical weather data at the local level contains many gaps, and the continuing collection of such data could be much improved. Strengthening the database of observed weather is critical to understanding the changes that have occurred already, to project future changes, and to plan appropriately to address them. Once collected and analyzed, climate data must be communicated in ways that help decision-makers understand climate impacts. Good tools are available—such as ClimateWizard.org and Servir ClimateServ—but practitioners at the local level must have the access and training to use them. Even in places where projections are uncertain, steps can be taken now to implement CSA practices and make farmers more resilient in the face of climate change.

Policy measures regarding adaptation to climate change include efforts to adjust socio-economic and ecologic systems. Colombia has undertaken various measures in terms of climate change mitigation and adaptation since becoming a party of the Kyoto protocol in 2001 and a party of the United Nations Framework Convention on Climate Change (UNFCCC) in 1995. The first national communication to the UNFCCC stated how Colombian agriculture will be severely impacted under different emission scenarios and time frames. The analyses in this document further support that climate change will severely threaten the socioeconomics of Colombian agriculture. We first query national data sources to characterize the agricultural sector. We then use 17 Global Circulation Model (GCM) outputs to quantify how Colombian agricultural production may be affected by climate change, and show the expected changes to years 2040–2069 (“2050”) under the A2 scenario of the Intergovernmental Panel on Climate Change Special Report on Emissions Scenarios (SRES-A2) and the overall trends in both precipitation and temperature to 2100. We then evaluate expected changes within different regions and measure the proportion of area affected within each crop’s distributional range. By 2050, climatic change in Colombia will likely impact 3.5 million people, 14 % of national GDP corresponding to agriculture, employment of 21 % of the population, agro-industries, supply chains, and food and nutritional security. If no adaptation measures are taken, 80 % of crops would be impacted in more than 60 % of their current areas of cultivation, with particularly severe impacts in high value perennial and exportable crops. Impacts also include soil degradation and organic matter losses in the Andes hillsides; likely flooding in the Caribbean and Pacific coasts; niche losses for coffee, fruit, cocoa, and bananas; changes in prevalence of pests and diseases; and increases in the vulnerabilities of non-technically developed smallholders. There is, however, still time to change the current levels of vulnerability if a multidisciplinary focus (i.e., agronomic, economic, and social) in vulnerable sectors is undertaken. Each sub-sector and the Government need to invest in: (1) data collection, (2) detailed, regionally-based impact assessments, (3) research and development, and (4) extension and technology transfer. Support to vulnerable smallholders should be given by the state in the form of agricultural insurance systems contextualized under the phenomenon of climate change. A national coordination scheme led by (but not restricted to) the Ministry of Agriculture and Rural Development (MADR) with the contributions of national and international institutions is needed to address agricultural adaptation.

This paper examines the impacts of climate change on cassava production in Africa, and questions whether cassava can play an important role in climate change adaptation. First, we examine the impacts that climate change will likely have on cassava itself, and on other important staple food crops for Africa including maize, millets, sorghum, banana, and beans based on projections to 2030. Results indicate that cassava is actually positively impacted in many areas of Africa, with −3.7% to +17.5% changes in climate suitability across the continent. Conversely, for other major food staples, we found that they are all projected to experience negative impacts, with the greatest impacts for beans (−16% ± 8.8), potato (−14.7 ± 8.2), banana (−2.5% ± 4.9), and sorghum (−2.66% ± 6.45). We then examined the likely challenges that cassava will face from pests and diseases through the use of ecological niche modeling for cassava mosaic disease, whitefly, brown streak disease and cassava mealybug. The findings show that the geographic distribution of these pests and diseases are projected to change, with both new areas opening up and areas where the pests and diseases are likely to leave or reduce in pressure. We finish the paper by looking at the abiotic traits of priority for crop adaptation for a 2030 world, showing that greater drought tolerance could bring some benefits in all areas of Africa, and that cold tolerance in Southern Africa will continue to be a constraint for cassava despite a warmer 2030 world, hence breeding needs to keep a focus on this trait. Importantly, heat tolerance was not found to be a major priority for crop improvement in cassava in the whole of Africa, but only in localized pockets of West Africa and the Sahel. The paper concludes that cassava is potentially highly resilient to future climatic changes and could provide Africa with options for adaptation whilst other major food staples face challenges.

Research into the origins of food plants has led to the recognition that specific geographical regions around the world have been of particular importance to the development of agricultural crops. Yet the relative contributions of these different regions in the context of current food systems have not been quantified. Here we determine the origins (‘primary regions of diversity’) of the crops comprising the food supplies and agricultural production of countries worldwide. We estimate the degree to which countries use crops from regions of diversity other than their own (‘foreign crops’), and quantify changes in this usage over the past 50 years. Countries are highly interconnected with regard to primary regions of diversity of the crops they cultivate and/or consume. Foreign crops are extensively used in food supplies (68.7% of national food supplies as a global mean are derived from foreign crops) and production systems (69.3% of crops grown are foreign). Foreign crop usage has increased significantly over the past 50 years, including in countries with high indigenous crop diversity. The results provide a novel perspective on the ongoing globalization of food systems worldwide, and bolster evidence for the importance of international collaboration on genetic resource conservation and exchange.

BACKGROUND: The author presents a 20-year experience leading cleft lip and palate surgical volunteer missions in Peru for CIRPLAST, a nonprofit volunteer plastic surgery goodwill program that has provided free surgery for patients with cleft lip and palate deformities in remote areas of Peru. Surgical procedures were performed by the author, together with a group of experienced plastic surgeons, under the auspices of the Peruvian Plastic Surgery Society, and local health authorities. METHODS: CIRPLAST missions are scheduled annually in different locations around Peru. Selected patients for surgery after adequate screening are photographed, and their cleft deformity is recorded. Scheduled patients or their parents, when they are minors, sign an informed consent form. Patients operated on in any given day are examined and photographed 1 day after surgery, before discharge. Between 30 and 35 patients are operated on at each mission site. About 2 weeks after the mission, patients are checked and photographed, and the outcome of surgery is recorded. Complications that may occur are recorded and treated by the CIRPLAST team as soon as possible. Almost all operations are performed under general endotracheal anesthesia coupled by local anesthesia containing a vasoconstrictor, to reduce bleeding and facilitate tissue dissection. All wounds of the lip and palate are closed with absorbable sutures, to avoid the need for suture removal. After cleft lip surgery, patients go to the recovery room for monitoring by nurses until they recover completely. RESULTS: A total of 6108 cleft lip and palate repairs, primary and secondary, were performed by CIRPLAST in 141 missions, between May 12, 1994, and October 15, 2014. The medical records of the 5162 patients (84.5%) who returned for follow-up (ranging from 12 days to 9 years) were reviewed retrospectively. Between 45% and 70% of the patients operated on a mission have returned for early follow-up and some the following year. There were 3176 males (51.9%) and 2932 females (48.1%). The incidence of isolated lip clefts was 1546 patients (25.3%); of isolated palate clefts, 2223 patients (36.4%); and combined defects, 2339 patients (38.3%). Of the 5162 patients who returned for follow-up, 377 patients (7.3%) had complications. Lip wound dehiscence was present in 58 patients (15.4). Palate fistula formation in 33 patients (8.8%): 24 (6.4%) after primary palate closure, and 9 (2.4%) after previous fistula closure. Infection occurred in 37 cleft lip patients (9.8%). Hypertrophic lip scars were seen in 56 patients (14.9%). Bleeding occurred in the recovery room after palatoplasty in 48 patients (12.7%), and in most cases, it was contained by applying pressure. No blood transfusions were used. Residual deformities of varying degree of the nose and/or lip occurred in 145 patients (38.5%). All required reoperation for correction. There were no intraoperative deaths in this series. CONCLUSIONS: During the past 20 years, the CIRPLAST team has offered free surgery with good outcomes and few complications, to more than 6000 cleft lip and/or palate patients in remote areas of Peru.

In immortalized GnRH neurons, cAMP production is elevated by increased extracellular Ca2+ and the Ca2+ channel agonist, BK-8644, and is diminished by low extracellular Ca2+ and treatment with nifedipine, consistent with the expression of adenylyl cyclase type I (AC I). Potassium-induced depolarization of GT1-7 neurons causes a dose-dependent monotonic increase in [Ca2+]i and elicits a bell-shaped cAMP response. The inhibitory phase of the cAMP response is prevented by pertussis toxin (PTX), consistent with the activation of G(i)-related proteins during depolarization. Agonist activation of the endogenous GnRH receptor in GT1-7 neurons also elicits a bell-shaped change in cAMP production. The inhibitory action of high GnRH concentrations is prevented by PTX, indicating coupling of the GnRH receptors to G(i)-related proteins. The stimulation of cAMP production by activation of endogenous LH receptors is enhanced by low (nanomolar) concentrations of GnRH but is abolished by micromolar concentrations of GnRH, again in a PTX-sensitive manner. These findings indicate that GnRH neuronal cAMP production is maintained by Ca2+ entry through voltage-sensitive calcium channels, leading to activation of Ca2+-stimulated AC I. Furthermore, the Ca2+ influx-dependent activation of AC I acts in conjunction with AC-regulatory G proteins to determine basal and agonist-stimulated levels of cAMP production.

The authors present their experience with 15 years of organizing cleft lip and palate surgical volunteer missions in Latin America. The history, basic principles, and objectives of Operation San Jose, a volunteer goodwill program from Christus St. Joseph Hospital in Houston, Texas, are covered. This report addresses the different problems encountered and solutions found. Following the principles set by Operation San Jose, CIRPLAST is a Peruvian foundation for plastic surgery that travels to remote areas in Peru, operating on patients with cleft lip and palate deformities. This report highlights the importance of working with local plastic surgeons and their residents, and emphasizes that the program should be organized by and the operations performed by accredited plastic surgeons and with the auspices and support of the national plastic surgery society and the local medical board. Operation San Jose promotes the creation of long-term benefits by offering a program to teach local surgeons cleft lip and palate repair techniques and to set up guidelines to organize local surgeons so that they can continue this effort by treating their own patients in their own countries.

The present study was designed to analyze the effects of glutamate (GLU) and its agonists on the release of noradrenaline (NA) from the mediobasal region of rat hypothalamus (MBH). Slices from hypothalamus were loaded in vitro with 3H-NA and thereafter exposed to GLU and the glutamate agonists N- methyl- d -aspartic acid (NMDA) and kainate (KA), in superfusion chambers. GLU evoked a significant 3H-NA release in a concentration-dependent manner. The EC50 was 35 mM. 6-Cyano-7-nitro-quinoxaline-2,3-dione (CNQX), a non-NMDA selective antagonist, and amino-7-phosphonoheptanoic acid (AP 7), a NMDA selective antagonist, both decreased the GLU-evoked response to about 50% of its value. NMDA, superfused in Mg2+-free Krebs-Ringer, exhibited a greater potency than GLU with an EC50 = 124 μM. KA was also able to evoke 3H-NA release, although overall responses to KA were lower than those of NMDA. The maximal response to KA was a 36% increase of release at a concentration of 200 μM. The effect of KA was blunted by CNQX. NMDA-induced 3H-NA release was progressively altered with age. In old rats (16–18 months) and middle-aged rats (10 months), responses to 200 μM NMDA were decreased respect to young (4 months) male rats. These results show that NMDA and KA receptors mediate the excitatory effects of GLU on NA release from nerve terminals in the MBH and suggest that GLU, in association with NA, participates in the complex mechanisms that regulate neuroendocrine functions. In addition, the age-related changes in NA responsiveness point to a GLU contribution on the maintenance of the reproductive status.

Histamine (HA) stimulates prolactin secretion via H1 and H2 receptors. In the present study, we examined the role of a third subtype of receptor recently described in brain, the H3-HA receptor, on prolactin secretion in male rats. R(-)alpha-methyl-HA (alpha-MHA), a selective H3 receptor agonist, was injected into the lateral ventricle of the brain in freely moving rats. alpha-MHA produced a dose-dependent (1-5 micrograms) and long-lasting increase in plasma prolactin levels. This increase was observed from 15 to 60 min after injection of alpha-MHA. Its stimulatory action was prevented by thioperamide (20 micrograms i.v.t), a selective H3 antagonist. This compound, injected intraventricularly, lacked effect by itself on basal plasma prolactin levels. Neither pyrilamine (H1 antagonist; 60 micrograms i.v.t.) nor ranitidine (H2 antagonist; 60 micrograms i.v.t.) affected alpha-MHA-induced prolactin release. The stimulatory effect was still present when brain HA was depleted by alpha-fluoromethylhistidine (30 mg/kg i.p.). Our findings suggest that alpha-MHA evokes prolactin release by activation of postsynaptic H3 receptors.

Serotonin (5-HT), the endogenous nonselective 5-HT receptor agonist, activates the inositol 1,4,5-triphosphate/calcium (InsP3/Ca2+) signaling pathway and exerts both stimulatory and inhibitory actions on cAMP production and GnRH release in immortalized GnRH neurons. The high degree of similarity between the signaling and secretory responses elicited by GnRH and 5-HT prompted us to target specific 5-HT receptor subtypes to deconvolute the complex actions of these agonists on signal transduction and GnRH release. Specific mRNA transcripts for 5-HT1A, 5-HT2C, 5-HT4, and 5-HT7 were identified in immortalized GnRH neurons (GT1-7). The rate of firing of spontaneous action potentials (APs) by hypothalamic GnRH neurons and cAMP production and pulsatile GnRH release in GT17 cells were profoundly inhibited during activation of the Gi-coupled 5-HT1A receptor. Treatment with a selective agonist to activate the Gq-coupled 5-HT2C receptor increased the rate of firing of spontaneous APs, stimulated InsP3 production and caused a delayed increase in GnRH release. Selective activation of the Gs-coupled 5-HT4 receptor also increased the rate of firing of APs, stimulated cAMP production, and caused a sustained and robust increase in GnRH release. The ability of 5-HT receptor subtypes expressed in GnRH neurons to activate single or multiple G proteins in a time- and dose-dependent manner differentially regulates the phospholipase C/InsP3/Ca2+, and adenylyl cyclase/cAMP signaling pathways, and thereby regulates the frequency and amplitude of pulsatile GnRH release. This process, in conjunction with the modulation of spontaneous electrical activity of the GnRH neuron, contributes to the control of the pulsatile mode of neuropeptide secretion that is characteristic of GnRH neuronal function in vivo and in vitro.

Immortalized GnRH neurons (GT1-7) express receptors for estrogen [estrogen receptor-alpha and -beta(ERalpha and ERbeta)] and progesterone (progesterone receptor A) and exhibit positive immunostaining for both intracellular and plasma membrane ERs. Exposure of GT1-7 cells to picomolar estradiol concentrations for 5-60 min caused rapid, sustained, and dose-dependent inhibition of cAMP production. In contrast, treatment with nanomolar estradiol concentrations for 60 min increased cAMP production. The inhibitory and stimulatory actions of estradiol on cAMP formation were abolished by the ER antagonist, ICI 182,780. The estradiol-induced inhibition of cAMP production was prevented by treatment with pertussis toxin, consistent with coupling of the plasma membrane ER to an inhibitory G protein. Coimmunoprecipitation studies demonstrated an estradiol-regulated stimulatory interaction between ERalpha and Galphai3 that was prevented by the ER antagonist, ICI 182,780. Exposure of perifused GT1-7 cells and hypothalamic neurons to picomolar estradiol levels increased the GnRH peak interval, shortened peak duration, and increased peak amplitude. These findings indicate that occupancy of the plasma membrane-associated ERs expressed in GT1-7 neurons by physiological estradiol levels causes activation of a Gi protein and modulates cAMP signaling and neuropeptide secretion.

The pulsatile secretion of gonadotropin-releasing hormone (GnRH) from normal and immortalized hypothalamic GnRH neurons is highly calcium-dependent and is stimulated by cAMP. It is also influenced by agonist activation of the endogenous GnRH receptor (GnRH-R), which couples to G q/11 as indicated by release of membrane-bound α q/11 subunits and increased inositol phosphate/Ca 2+ signaling. Conversely, GnRH antagonists increase membrane-associated α q/11 subunits and abolish pulsatile GnRH secretion. GnRH also stimulates cAMP production but at high concentrations has a pertussis toxin-sensitive inhibitory effect, indicative of receptor coupling to G i . Coupling of the agonist-activated GnRH-R to both G s and G i proteins was demonstrated by the ability of nanomolar GnRH concentrations to reduce membrane-associated α s and α i3 levels and of higher concentrations to diminish α i3 levels. Conversely, α i3 was increased during GnRH antagonist and pertussis toxin treatment, with concomitant loss of pulsatile GnRH secretion. In cholera toxin-treated GnRH neurons, decreases in α s immunoreactivity and increases in cAMP production paralleled the responses to nanomolar GnRH concentrations. Treatment with cholera toxin and 8-bromo-cAMP amplified episodic GnRH pulses but did not affect their frequency. These findings suggest that an agonist concentration-dependent switch in coupling of the GnRH-R between specific G proteins modulates neuronal Ca 2+ signaling via G s -cAMP stimulatory and G i -cAMP inhibitory mechanisms. Activation of G i may also inhibit GnRH neuronal function and episodic secretion by regulating membrane ion currents. This autocrine mechanism could serve as a timer to determine the frequency of pulsatile GnRH release by regulating Ca 2+ - and cAMP-dependent signaling and GnRH neuronal firing.

Episodic hormone secretion is a characteristic feature of the hypothalamo-pituitary-gonadal system, in which the profile of gonadotropin release from pituitary gonadotrophs reflects the pulsatile secretory activity of GnRH-producing neurons in the hypothalamus. Pulsatile release of GnRH is also evident in vitro during perifusion of immortalized GnRH neurons (GT1–7 cells) and cultured fetal hypothalamic cells, which continue to produce bioactive GnRH for up to 2 months. Such cultures, as well as hypothalamic tissue from adult rats, express GnRH receptors as evidenced by the presence of high-affinity GnRH binding sites and GnRH receptor transcripts. Furthermore, individual GnRH neurons coexpress GnRH and GnRH receptors as revealed by double immunostaining of hypothalamic cultures. In static cultures of hypothalamic neurons and GT1–7 cells, treatment with the GnRH receptor antagonist, [d-pGlu1, d-Phe2, d-Trp3,6]GnRH caused a prominent increase in GnRH release. In perifused hypothalamic cells and GT1–7 cells, treatment with the GnRH receptor agonist, des-Gly10-[d-Ala6]GnRH N-ethylamide, reduced the frequency and increased the amplitude of pulsatile GnRH release, as previously observed in GT1–7 cells. In contrast, exposure to the GnRH antagonist analogs abolished pulsatile secretion and caused a sustained and progressive increase in GnRH release. These findings have demonstrated that GnRH receptors are expressed in hypothalamic GnRH neurons, and that receptor activation is required for pulsatile GnRH release in vitro. The effects of GnRH agonist and antagonist analogs on neuropeptide release are consistent with the operation of an ultrashort-loop autocrine feedback mechanism that exerts both positive and negative actions that are necessary for the integrated control of GnRH secretion from the hypothalamus.

Agonist activation of cholinergic receptors expressed in perifused hypothalamic and immortalized GnRH-producing (GT1-7) cells induced prominent peaks in GnRH release, each followed by a rapid decrease, a transient plateau, and a decline to below basal levels. The complex profile of GnRH release suggested that acetylcholine (ACh) acts through different cholinergic receptor subtypes to exert stimulatory and inhibitory effects on GnRH release. Whereas activation of nicotinic receptors caused a transient increase in GnRH release, activation of muscarinic receptors inhibited basal GnRH release. Nanomolar concentrations of ACh caused dose-dependent inhibition of cAMP production that was prevented by pertussis toxin (PTX), consistent with the activation of a plasma-membrane Gi protein. Micromolar concentrations of ACh also caused an increase in phosphoinositide hydrolysis that was inhibited by the M1 receptor antagonist, pirenzepine. In ACh-treated cells, immunoblot analysis revealed that membrane-associated G(alpha q/11) immunoreactivity was decreased after 5 min but was restored at later times. In contrast, immunoreactive G(alpha i3) was decreased for up to 120 min after ACh treatment. The agonist-induced changes in G protein alpha-subunits liberated during activation of muscarinic receptors were correlated with regulation of their respective transduction pathways. These results indicate that ACh modulates GnRH release from hypothalamic neurons through both M1 and M2 muscarinic receptors. These receptor subtypes are coupled to Gq and Gi proteins that respectively influence the activities of PLC and adenylyl cyclase/ion channels, with consequent effects on neurosecretion.