Role of p38MAPK in ultrastructural alterations of desmosomes in human ex vivo pemphigus model

Desmosomes interconnect epithelial cells together and are abundant in tissues constantly challenged by shear forces. They are composed of different isoforms of intercellular adhesion proteins which include desmogleins (Dsg) and desmocollins (Dsc). These proteins of apposing cells interact in homophilic and heterophilic manner thereby conferring integrity to the tissue. When this crucial role of desmosomes is compromised, several desmosome-associated diseases such as pemphigus may occur. Pemphigus is a blistering disease of the skin and oral mucosa. It is caused by anti-Dg3 and anti-Dsg1 autoantibodies that bind to the extracellular domains of the desmogleins and perturb their interaction. There are different phenotypes of the disease depending on the autoantibody profiles. Pemphigus vulgaris (PV) is caused by autoantibodies (PV-IgG) targeting Dsg1 and Dsg3 whereas pemphigus foliaceus (PF) is associated with autoantibodies against Dsg1 only. Pemphigus vulgaris is recognized as two sub-types; i.e, the mucosal-dominant form (mdPV) caused by anti-Dsg3 autoantibodies and the mucocutaneus (mcPV) variant caused by both anti-Dsg1 and anti-Dsg3 autoantibodies. Several lines of evidence demonstrated that pemphigus is caused by disruption of Dsg interaction when the autoantibodies are interposed between the interacting Dsg (steric hindrance) and signaling triggered by autoantibody binding. It has been widely accepted that different signaling pathways work in concert in the modulation of desmosome structure and dynamics. p38 mitogen activated protein kinase (p38MAPK) has been extensively studied and its phosphorylation was detected in cell cultures in response to PV-IgG binding as well as in skin lesions of pemphigus patients. Pharmacological inactivation of this pathway attenuated cell dissociation in cultures and blister formation in murine models. However, no data was available with respect to the role of p38MAPK in blister formation in human skin and mucosa. Therefore, we tested the dependency of blister formation and desmosome ultrastructural alteration on p38MAPK signaling induced by PV-IgG in human skin and mucosa explant cultures. Accordingly, we adapted the existing ex vivo skin model and also established a novel ex vivo mucosa model, and employed histological, immune-histochemical as well as electron microscopy analyses to determine the role of p38MAPK signaling in PV pathogenesis. Human skin biopsies were treated with the mouse monoclonal Dsg3-specific antibodyAK23, in comparison to antibody fractions from patients with mucocutaneous PV (mcPV-IgG) or mucosal PV (mdPV-IgG). mcPV-IgG only were sufficient to induce blisters as well as alterations in desmosome ultrastructure. In contrast, in human labial mucosa explants both AK23 and mdPV-IgG were sufficient to induce blisters as well as alterations in desmosome ultrastructure. Moreover, inhibition of p38MAPK using the specific inhibitor SB202190 was effective to avert blister formation, rescue desmosome size and number as well as preserved keratin filament association with desmosomal plaques in human skin. However, in the newly established human ex vivo mucosa model, inhibition of p38MAPK with specific inhibitors SB202190 and SB203580 was not effective to prevent these alterations. Taken together, our data demonstrate that p38MAPK plays a key role in blister formation through modulation of desmosome ultrastructure in human skin. In contrast, blister formation and associated ultrastructural changes of desmosomes in mucosa may depend on steric hindrance and other signaling pathways independent of p38MAPK

Role of p38MAPK in ultrastructural alterations of desmosomes in human ex vivo pemphigus model

Desmosomes interconnect epithelial cells together and are abundant in tissues constantly challenged by shear forces. They are composed of different isoforms of intercellular adhesion proteins which include desmogleins (Dsg) and desmocollins (Dsc). These proteins of apposing cells interact in homophilic and heterophilic manner thereby conferring integrity to the tissue. When this crucial role of desmosomes is compromised, several desmosome-associated diseases such as pemphigus may occur. Pemphigus is a blistering disease of the skin and oral mucosa. It is caused by anti-Dg3 and anti-Dsg1 autoantibodies that bind to the extracellular domains of the desmogleins and perturb their interaction. There are different phenotypes of the disease depending on the autoantibody profiles. Pemphigus vulgaris (PV) is caused by autoantibodies (PV-IgG) targeting Dsg1 and Dsg3 whereas pemphigus foliaceus (PF) is associated with autoantibodies against Dsg1 only. Pemphigus vulgaris is recognized as two sub-types; i.e, the mucosal-dominant form (mdPV) caused by anti-Dsg3 autoantibodies and the mucocutaneus (mcPV) variant caused by both anti-Dsg1 and anti-Dsg3 autoantibodies. Several lines of evidence demonstrated that pemphigus is caused by disruption of Dsg interaction when the autoantibodies are interposed between the interacting Dsg (steric hindrance) and signaling triggered by autoantibody binding. It has been widely accepted that different signaling pathways work in concert in the modulation of desmosome structure and dynamics. p38 mitogen activated protein kinase (p38MAPK) has been extensively studied and its phosphorylation was detected in cell cultures in response to PV-IgG binding as well as in skin lesions of pemphigus patients. Pharmacological inactivation of this pathway attenuated cell dissociation in cultures and blister formation in murine models. However, no data was available with respect to the role of p38MAPK in blister formation in human skin and mucosa. Therefore, we tested the dependency of blister formation and desmosome ultrastructural alteration on p38MAPK signaling induced by PV-IgG in human skin and mucosa explant cultures. Accordingly, we adapted the existing ex vivo skin model and also established a novel ex vivo mucosa model, and employed histological, immune-histochemical as well as electron microscopy analyses to determine the role of p38MAPK signaling in PV pathogenesis. Human skin biopsies were treated with the mouse monoclonal Dsg3-specific antibodyAK23, in comparison to antibody fractions from patients with mucocutaneous PV (mcPV-IgG) or mucosal PV (mdPV-IgG). mcPV-IgG only were sufficient to induce blisters as well as alterations in desmosome ultrastructure. In contrast, in human labial mucosa explants both AK23 and mdPV-IgG were sufficient to induce blisters as well as alterations in desmosome ultrastructure. Moreover, inhibition of p38MAPK using the specific inhibitor SB202190 was effective to avert blister formation, rescue desmosome size and number as well as preserved keratin filament association with desmosomal plaques in human skin. However, in the newly established human ex vivo mucosa model, inhibition of p38MAPK with specific inhibitors SB202190 and SB203580 was not effective to prevent these alterations. Taken together, our data demonstrate that p38MAPK plays a key role in blister formation through modulation of desmosome ultrastructure in human skin. In contrast, blister formation and associated ultrastructural changes of desmosomes in mucosa may depend on steric hindrance and other signaling pathways independent of p38MAPK

Haematological and Serum Biochemical Parameters of Mature Harco Cocks Treated with Human Menopausal Gonadotrophin (Diclair) For Spermatogenesis

Twenty sexually matured (24 weeks old) healthy Harco cocks were used to determine the effect of Gonadotrophin (Diclair®) on haematology and serum biochemistry. The cocks were divided into 4 treatment groups of 5 cocks per group identified as T1 (control) administered with 1ml physiological saline, T2, administered with 6.75i.u Diclair® and T4, administered with 20.25i.u Diclair®, with one cock per replicate in a completely Randomized Design (CRD). The injections were dividedinto three doses each and administered intramuscularly in the thigh for three consecutive days. One week after Diclair® treatments, five birds from each group were bled from the wing veins for haematology and serum biochemistry. Results of this study showed significant differences (P0.05) among the treatment groups. Basophils were not detected among the treatment groups. The results further showed significant differences (P0.05) among the treatment groups. However, the values were within the normal ranges, indicating that Diclair® had no deleterious effect on these parameters

Kidney Function Test, Weight Gain and Serum Protein Values of Mature Male Turkeys Treated with Gonadotrophin (Diclair) For Sperm Production

Sixteen sexually matured (12 months old) healthy male Turkeys were used to determine the effect of Gonadotrophin (Diclair®) on kidney function, weight gain and serum protein values. The Turkeys were divided into 4 treatment groups, identified as T1 (control) administered with 1.00ml physiological saline (0.00 i.u Diclair®), T2 , administered with 13.50 i.u Diclair®, T3,administered with 27.00i.u Dicliar®T4, administered with 40.50 i.u Dicliar(R), with one Turkey per replicate in a completely Randomized Design (CRD). The injections were divided into 3 doses each and administered intramuscularly in the thigh for three consecutive days. Blood was collected one week after Diclair® administration. Four Turkeys were randomly selected fro-m each treatment groupand bled to collect blood for blood chemistry analysis. The Turkey were weighed every week for five weeks and their weight recorded. The result showed that there were significant differences (P 0.05) among the treatment groups. The results further showed that there were no significant differences (p > 0.05) among the treatment groups in initial body weight. However, there were significant differences (P< 0.05) among the treatment groups in final body weight and weight gain. Similarly there were significant differences (P< 0.05) among the treatment groups in all the serum protein values measure: albumin, globulin, serum total protein as well as albumin/globulin ratio. The results of the study showed that Diclair enhanced kidney function and weight gain without any deleterious effects on serum protein values of the male Turkeys

Re-creando el espacio publico urbano. Politica para construir ciudad y ciudadania en Venezuela

En 1996 se crea, en la Alcaldía de Maracaibo, el Instituto Municipal de Ambiente (IMA) para hacer de Maracaibo una “ciudad bonita” y atraer inversiones, recuperando los espacios públicos urbanos, ámbitos clave para la comunicación, la cohesión social y la construcción de ciudad y ciudadanía. El artículo evalúa el impacto de esta política en la ordenación territorial-urbana e imagen de Maracaibo y en la formación de ciudadanía, durante el periodo 1996-2006. Se realizó una revisión documental y de campo, se identificaron y localizaron los espacios públicos recuperados, el modelo de gestión implantado y aplicaron cuestionarios para conocer la opinión de los ciudadanos. Concluyendo que, la recuperación del espacio publico ha impactado positivamente la estructura-imagen de Maracaibo, reconocida por residentes y visitantes y, la autoestima-ciudadanía del marabino y destacando la necesidad de actuar en tres frentes: rechazando los proyectos tendentes a la privatización del espacio público; promoviendo el mantenimiento de los espacios recuperados con la participación activa de los ciudadanos y la cultura de sostenibilidad y, estimulando la aplicación de esta política en los bordes de la ciudad -periferia urbana-, como vía hacia una ciudad y ciudadanía sostenible

Assessment of Forest Investment, Financial Flows and Revenue Collection in the Abia State Forest Sector, Nigeria

Varieties of resources abound in the forests of Nigeria and especially in Abia state, an economic treasure house of resources. Sustainable management of the forests in Nigeria is crucial for a consistent supply of forest resources. Ten forest reserves were surveyed to determine the extent of the government’s involvement in sustainable forest management in Abia State. The government consider the forest reserves, as a revenue-generating venture, leading to the excessive exploitation of forest resources in the state. The exploitation is without regard for sustainability as the harvest is consistently higher than its growth. The government’s involvement in funding the forestry sector over these years has&nbsp; been the payment of salaries to staff of the State forestry department and revenue collection, while international donors,&nbsp; corporate organizations and private sectors have not considerably invested in the Abia State forest sector. The survey&nbsp; revealed the challenges militating the productivity of the forest sector in Abia State, namely: inadequate capital,&nbsp; administrative incompetence and bureaucratic bottleneck, political instability, ecological challenge, insufficient skilled&nbsp; personnel, corruption, lack of forest equipment and inefficient forest laws. Training should be organized for the forest&nbsp; staff and communities to ensure the sustainable use of forest resources. There is a need for the modernization of forestry practice in Abia State and all that go with it being accorded a well-deserved priority in the present economic dispensation in Nigeria

Mechanisms Causing Acantholysis in Pemphigus-Lessons from Human Skin

Pemphigus vulgaris (PV) is an autoimmune bullous skin disease caused primarily by autoantibodies (PV-IgG) against the desmosomal adhesion proteins desmoglein (Dsg)1 and Dsg3. PV patient lesions are characterized by flaccid blisters and ultrastructurally by defined hallmarks including a reduction in desmosome number and size, formation of split desmosomes, as well as uncoupling of keratin filaments from desmosomes. The pathophysiology underlying the disease is known to involve several intracellular signaling pathways downstream of PV-IgG binding. Here, we summarize our studies in which we used transmission electron microscopy to characterize the roles of signaling pathways in the pathogenic effects of PV-IgG on desmosome ultrastructure in a human ex vivo skin model. Blister scores revealed inhibition of p38MAPK, ERK and PLC/Ca2+ to be protective in human epidermis. In contrast, inhibition of Src and PKC, which were shown to be protective in cell cultures and murine models, was not effective for human skin explants. The ultrastructural analysis revealed that for preventing skin blistering at least desmosome number (as modulated by ERK) or keratin filament insertion (as modulated by PLC/Ca2+) need to be ameliorated. Other pathways such as p38MAPK regulate desmosome number, size, and keratin insertion indicating that they control desmosome assembly and disassembly on different levels. Taken together, studies in human skin delineate target mechanisms for the treatment of pemphigus patients. In addition, ultrastructural analysis supports defining the specific role of a given signaling molecule in desmosome turnover at ultrastructural level

The Impact of South Africa’s Total Early-stage Entrepreneurial Activity Rate on Entrepreneurial Attitudes and Behavior

Given that entrepreneurship scholars use various approaches to measure entrepreneurial success, which leads to inconsistent findings, in this study we investigate the impact that South Africa’s total early-stage entrepreneurial activity (TESEA) rate has on the entrepreneurial attitudes and behavior of South Africans and their reactions to different macroeconomic factors. Secondary datasets were elicited from the World Bank’s world development indicators and the Global Entrepreneurship Monitor (GEM) database from 2003–2014. Using a generalized linear model (GLM) poisson regression, we affirmed the statistical significance of these relationships, as well as enhanced the validity and reliability of the quantitative estimation procedure. We found that the TESEA rate is positively associated with a higher ease of doing business and entrepreneurial intention level. Furthermore, we found that when entrepreneurship is viewed as a good career choice, and given media visibility, the TESEA rate increases, and vice versa. It was also observed that the GDP per capita (which relies on efficiency, innovation and optimal utilization of market knowledge), as well as the level of economic freedom in South Africa, positively impacts the nation’s TESEA rate. Likewise, we found that the unemployment rate is positively associated with the TESEA rate of South Africa. South Africa’s TESEA rate is influenced by a combination of factors due to the dynamic nature of individual traits. However, the TESEA rate has a remarkable impact on South Africa’s entrepreneurial decision making. This implies that a targeted approach is more relevant when implementing both private and government policy initiatives

Implementation of automatic frequency control system at samanalawewa power station

Electricity cannot be stored, what is produced has to be consumed immediately. The frequency control theory is used for balancing the power system; it means electrical energy should always be equal to summation of demand and system losses. Samanalawewa Power Station where the frequency control system is operated manually has experienced the need of an automatic frequency controlling system. Therefore a microcontroller based automatic frequency control unit is used to correct the errors mainly due to human factors and also it is more accurate than manual system and gives lot of benefits. Finally it saves the valuable time and money to the public. Therefore automatic frequency control system is essential. The system basically composes with a microcontroller, an LCD Display, Relays and a Selector switch. This report describes the theoretical background on frequency control, concepts and implementation, about the microcontroller, preparation, software installation and programming, Result interpretation and finally the conclusion of automatic frequency control to be implemented at the Samanalawewa Power Station

Role of PKC and ERK Signaling in Epidermal Blistering and Desmosome Regulation in Pemphigus

Desmosomes reinforce cohesion of epithelial cells at the interface between adjacent cells. They include the cadherin-type adhesion molecules desmoglein 1 (Dsg1) and Dsg3. Pemphigus vulgaris (PV) is an autoimmune disease in which circulating autoantibodies (PV-IgG) targeting Dsg1 and 3 cause characteristic epidermal blister formation. It has been shown that PV-IgG binding induced activation of kinases such as ERK and PKC, and inhibition of these signaling pathways prevented loss of cell cohesion in cell cultures. However, the role of Erk and PKC in blister formation and regulation of desmosome ultrastructure in human skin are unknown. Accordingly, we assessed the role of PKC and ERK signaling pathways in blister formation and regulation of desmosome ultrastructure in human epidermis. Here we performed electron microscopy analyses using human skin explants injected with PV-IgG together with inhibitors for PKC or ERK signaling. Inhibition of PKC was not effective to prevent suprabasal blister formation or ultrastructural alterations of desmosomes. In contrast, inhibition of ERK signaling significantly ameliorated blister formation and decrease in the number of desmosomes whereas shortening and splitting of desmosomes and keratin filament insertion were not different from samples treated with PV-IgG alone. However, apical desmosomes between basal and suprabasal cells remained unaltered when ERK signaling was inhibited. Therefore, our results show that inhibition of ERK but not PKC signaling appears to be effective to ameliorate blistering and alterations of desmosome ultrastructure triggered by PV-IgG in human skin