G then able to bind inner PM phospholipids as well as

G then able to bind inner PM phospholipids as well as cytoplasmic membranes of organelles (Fig. 3d; Table 1); and/or (ii) incubated with cells to target outer leaflet phospholipids after transbilayer flip-flop. The pleckstrin homology (PH) domain is one of these well-characterized probes specific for phosphoinositides (PIs; [122]). The 100 amino acid-PH domain is contained in several proteins, such as pleckstrin or phospholipase C (PLC), with distinct binding affinity for GSK-AHAB site different PIs [123]. For instance, PH domain of PLC (PH-PLC) has a high affinity for phosphatidylinositol-4,5-bisphosphate (PIP2) [124, 125]. The discoidin C2 domain is another probe, specific for phosphatidylserine (PS). The 160 amino acid-discoidin C2 domain is present in blood coagulation factors V and VIII, milk fat globule-EGF factor 8 (MFGE8; also known as lactadherin [Lact-C2]) and other plasma proteins. PH or discoidin C2 domains can be fluorescently tagged, allowing to study phospholipid membrane distribution [126-128]. Other globular domains capable to bind phospholipids at the membrane surface include: (i) the FYVE zinc finger domain found in EEA1 (Early Endosome Antigen 1) a.o. that binds to phosphatidylinositol-3-phosphate (PI3P); and (ii) the calcium-dependent phospholipid binding Annexins, such as Annexin A2, which preferentially interacts with PIP2, or Annexin A5, which is currently the most commonly used probe for PS targeting at outer PM leaflet [129]. To further overcome limitation due to lack of PS labeling at the luminal membrane leaflet of organelles. Parton and coll. recently developed a novel on-section labeling approach on fast-frozen sample using purified GST (glutathione-S-transferase)-Lact-C2 fusion protein followed by transmission electron microscopy. This technique is based on PM01183 site high-pressure freezing, freeze-substitution with minimal fixatives and embedding at low temperature. Sections are then fixed, labeled with purified GST-Lact-C2 and followed by detection with anti-GST antibody and protein A?Author Manuscript Author Manuscript Author Manuscript Author ManuscriptProg Lipid Res. Author manuscript; available in PMC 2017 April 01.Carquin et al.Pagegold. Such method avoids cell permeabilization as well as detergent extraction [126]. For more details on phospholipid-binding domains, please refer to [130]. Similarly to other probes, this approach also presents limitations including perturbation of normal lipid function upon high expression and high variability of affinity and specificity [129, 131]. 3.1.3. Antibodies, Fab fragments and nanobodies–Antibodies have been recognized as gold standard to detect proteins. Interestingly, several antibodies have also been generated to decorate PM lipids (Fig. 3e). For example, there are monoclonal antibodies (mAbs) produced to detect specific GSLs expressed during the differentiation of oligodendrocytes and used for studying their in vitro maturation: (i) the mAb A2B5, against gangliosides GD3, GT3 and O-acetylated GT3 in early oligodendrocyte progenitors; (ii) the mAb O4, against sulfated GSLs expressed by late progenitors; and (iii) the mAb O1 and the mAb Ranscht, against galactosylceramides in mature oligodendrocytes (for a review, see [132]). These antibodies have revealed submicrometric GSL-enriched domains at different stages of oligodendrocyte differentiation, as illustrated in Table 1. Although less developed, antibodies are also used to decorate phospholipids. For example, the role of PS do.G then able to bind inner PM phospholipids as well as cytoplasmic membranes of organelles (Fig. 3d; Table 1); and/or (ii) incubated with cells to target outer leaflet phospholipids after transbilayer flip-flop. The pleckstrin homology (PH) domain is one of these well-characterized probes specific for phosphoinositides (PIs; [122]). The 100 amino acid-PH domain is contained in several proteins, such as pleckstrin or phospholipase C (PLC), with distinct binding affinity for different PIs [123]. For instance, PH domain of PLC (PH-PLC) has a high affinity for phosphatidylinositol-4,5-bisphosphate (PIP2) [124, 125]. The discoidin C2 domain is another probe, specific for phosphatidylserine (PS). The 160 amino acid-discoidin C2 domain is present in blood coagulation factors V and VIII, milk fat globule-EGF factor 8 (MFGE8; also known as lactadherin [Lact-C2]) and other plasma proteins. PH or discoidin C2 domains can be fluorescently tagged, allowing to study phospholipid membrane distribution [126-128]. Other globular domains capable to bind phospholipids at the membrane surface include: (i) the FYVE zinc finger domain found in EEA1 (Early Endosome Antigen 1) a.o. that binds to phosphatidylinositol-3-phosphate (PI3P); and (ii) the calcium-dependent phospholipid binding Annexins, such as Annexin A2, which preferentially interacts with PIP2, or Annexin A5, which is currently the most commonly used probe for PS targeting at outer PM leaflet [129]. To further overcome limitation due to lack of PS labeling at the luminal membrane leaflet of organelles. Parton and coll. recently developed a novel on-section labeling approach on fast-frozen sample using purified GST (glutathione-S-transferase)-Lact-C2 fusion protein followed by transmission electron microscopy. This technique is based on high-pressure freezing, freeze-substitution with minimal fixatives and embedding at low temperature. Sections are then fixed, labeled with purified GST-Lact-C2 and followed by detection with anti-GST antibody and protein A?Author Manuscript Author Manuscript Author Manuscript Author ManuscriptProg Lipid Res. Author manuscript; available in PMC 2017 April 01.Carquin et al.Pagegold. Such method avoids cell permeabilization as well as detergent extraction [126]. For more details on phospholipid-binding domains, please refer to [130]. Similarly to other probes, this approach also presents limitations including perturbation of normal lipid function upon high expression and high variability of affinity and specificity [129, 131]. 3.1.3. Antibodies, Fab fragments and nanobodies–Antibodies have been recognized as gold standard to detect proteins. Interestingly, several antibodies have also been generated to decorate PM lipids (Fig. 3e). For example, there are monoclonal antibodies (mAbs) produced to detect specific GSLs expressed during the differentiation of oligodendrocytes and used for studying their in vitro maturation: (i) the mAb A2B5, against gangliosides GD3, GT3 and O-acetylated GT3 in early oligodendrocyte progenitors; (ii) the mAb O4, against sulfated GSLs expressed by late progenitors; and (iii) the mAb O1 and the mAb Ranscht, against galactosylceramides in mature oligodendrocytes (for a review, see [132]). These antibodies have revealed submicrometric GSL-enriched domains at different stages of oligodendrocyte differentiation, as illustrated in Table 1. Although less developed, antibodies are also used to decorate phospholipids. For example, the role of PS do.

Them cope with their losses. Not only is this a strengths-based

Them cope with their losses. Not only is this a strengths-based approach (McGovern, 2011), but the interaction helps each couple move beyond the current situation and look at it in the context of their whole sharedDementia (London). Author manuscript; available in PMC 2016 July 01.Ingersoll-Dayton et al.Pagelife together, recognizing the buy Procyanidin B1 individuality and fullness of their lives, transcending some of the roles they have assumed because of the illness. The intervention addresses them as a couple working as partners in the context of a long partnership, instead of limiting them to the roles of caregiver and care receiver. It helps them to integrate their experiences, remember high points and low points and, most importantly, relive them together. It solidifies their relationship and their identity as a couple with a long history. We found that in both the United States and Japan, this dyadic approach brought the person with dementia into the conversation. People with dementia, or even early memory loss, are often excluded from this kind of conversation or talked to in a condescending manner (Hamaguchi, 2011). The modeling and encouragement to talk that the interventionists gave to the person with dementia helped the partner learn ways of encouraging their spouse with memory loss to participate. This approach helped to normalize the dementia experience and move away from the perception of the person with dementia as a victim. Taken together, our experiences with the Couples Life Story Approach suggest that it is a promising dyadic model that can be easily translated across cultures. The American and Japanese practitioners found the intervention easy to implement and adaptable to their personal styles as well. While the kinds of couples seen in Japan and the United States have been somewhat different, these Sodium lasalocid supplier variations have helped us feel confident that the Couples Life Story Approach is applicable to many kinds of couples. We welcome other practitioners working in dementia care to use and adapt the Couples Life Story Approach to their own cultural contexts.Author Manuscript Author Manuscript Author Manuscript Author ManuscriptBiographiesBerit Ingersoll-Dayton is a social worker and a social psychologist. Her research focuses on social relationships in later life, including cross-cultural similarities and differences. She is a Professor in the School of Social Work at the University of Michigan, USA where she is Principal Investigator of the Couples Life Story Project. Beth Spencer is a geriatric social worker specializing in dementia care. Her clinical and research interests focus on caregivers and individuals with memory loss. She is a Project Manager for the Hartford Center of Excellence in Geriatric Social Work at the University of Michigan, USA and also Co-Investigator of the Couples Life Story Project. Ruth Campbell is a social worker specializing in gerontology. Her areas of interest are caregiving and dementia in the United States and Japan, changing family relationships in Japan, and the national long-term care insurance system in Japan. Retired from the University of Michigan where she was Associate Director for Social Work and Community Programs in the Geriatrics Center, she is now affiliated with Keiseikai Gerontology Institute in Tokyo, Japan. Yukiko Kurokawa is a clinical psychologist. Her research focuses on psychotherapy and other interventions for older adults and their families. She is a Professor in the School of Psycholog.Them cope with their losses. Not only is this a strengths-based approach (McGovern, 2011), but the interaction helps each couple move beyond the current situation and look at it in the context of their whole sharedDementia (London). Author manuscript; available in PMC 2016 July 01.Ingersoll-Dayton et al.Pagelife together, recognizing the individuality and fullness of their lives, transcending some of the roles they have assumed because of the illness. The intervention addresses them as a couple working as partners in the context of a long partnership, instead of limiting them to the roles of caregiver and care receiver. It helps them to integrate their experiences, remember high points and low points and, most importantly, relive them together. It solidifies their relationship and their identity as a couple with a long history. We found that in both the United States and Japan, this dyadic approach brought the person with dementia into the conversation. People with dementia, or even early memory loss, are often excluded from this kind of conversation or talked to in a condescending manner (Hamaguchi, 2011). The modeling and encouragement to talk that the interventionists gave to the person with dementia helped the partner learn ways of encouraging their spouse with memory loss to participate. This approach helped to normalize the dementia experience and move away from the perception of the person with dementia as a victim. Taken together, our experiences with the Couples Life Story Approach suggest that it is a promising dyadic model that can be easily translated across cultures. The American and Japanese practitioners found the intervention easy to implement and adaptable to their personal styles as well. While the kinds of couples seen in Japan and the United States have been somewhat different, these variations have helped us feel confident that the Couples Life Story Approach is applicable to many kinds of couples. We welcome other practitioners working in dementia care to use and adapt the Couples Life Story Approach to their own cultural contexts.Author Manuscript Author Manuscript Author Manuscript Author ManuscriptBiographiesBerit Ingersoll-Dayton is a social worker and a social psychologist. Her research focuses on social relationships in later life, including cross-cultural similarities and differences. She is a Professor in the School of Social Work at the University of Michigan, USA where she is Principal Investigator of the Couples Life Story Project. Beth Spencer is a geriatric social worker specializing in dementia care. Her clinical and research interests focus on caregivers and individuals with memory loss. She is a Project Manager for the Hartford Center of Excellence in Geriatric Social Work at the University of Michigan, USA and also Co-Investigator of the Couples Life Story Project. Ruth Campbell is a social worker specializing in gerontology. Her areas of interest are caregiving and dementia in the United States and Japan, changing family relationships in Japan, and the national long-term care insurance system in Japan. Retired from the University of Michigan where she was Associate Director for Social Work and Community Programs in the Geriatrics Center, she is now affiliated with Keiseikai Gerontology Institute in Tokyo, Japan. Yukiko Kurokawa is a clinical psychologist. Her research focuses on psychotherapy and other interventions for older adults and their families. She is a Professor in the School of Psycholog.

A into oligomers and has a clearance effect on the existing

A into oligomers and has a clearance effect on the existing A (Cole et al. 2007). A very interesting in vivo approach with multiphoton microscopy showed the VelpatasvirMedChemExpress Velpatasvir ability of curcumin to cross the blood-brain barrier (BBB) and disrupt amyloid plaques (GarciaAlloza et al. 2007). Interestingly, curcumin possesses both MAO-A- and MAO-B-inhibiting properties and has been shown to modulate the levels of noradrenaline, dopamine and serotonin in the brain, demonstrating antidepressant effects in animal models of depression (Scapagnini et al. 2012) and in patients with major depressive disorder (Sanmukhani et al. 2013). Much of the research conducted to date on curcumin has been focused on exploring its protective and therapeutic effects against age-related degeneration. Recently the possibility that curcumin and its metabolites can modulate pathways directly involved in the determination of lifespan and extension of longevity, has been also highlighted (Shen et al. 2013). Tetrahydrocurcumin (THC), an active metabolite of curcumin, produced after its ingestion, has been shown to extend lifespan of drosophila under normal conditions, by attenuating oxidative stress via FOXO and Sir2 modulation (Xiang et al. 2011). Curcuminoids may also affect mammalian longevity, as shown in mice fed diets containing THC starting at the age of 13 months, which showed significantly increased mean lifespan (Shen et al. 2013).Author Manuscript Author Manuscript Author Manuscript Author 1-Deoxynojirimycin price ManuscriptSummary and ConclusionsThe traditional diet in Okinawa is based on green and yellow vegetables, root vegetables (principally sweet potatoes), soybean-based foods, and other plants, many with medicinal properties. This is supplemented by regular seafood consumption and consumption of smaller amounts of lean meats, fruit, and medicinal garnishes and spices. Sanpin (jasmine) tea is the principal beverage, consumed with meals and awamori (Okinawan sake) is the social drink of choice. The dietary composition over the past half-century has changed from a low calorie diet dominated by low glycemic index carbohydrates, low in protein and fat, to one more moderate in all three macronutrients. While the caloric content has increased due to higherMech Ageing Dev. Author manuscript; available in PMC 2017 April 24.Willcox et al.Pageconsumption of calorically dense foods, the diet remains very healthy by most expert criteria including the National Cholesterol Education Program (NCEP), the U.S. Dietary Guidelines for Americans Advisory Committee, and the Unified Dietary Guidelines. Many of the characteristics of the traditional Okinawan diet are shared with other healthy dietary patterns, such as the traditional Mediterranean diet, the modern DASH diet, and the modern Portfolio diet. All these dietary patterns have been found to be associated with reduced risk for cardiovascular disease (Appel, 2008; Fung et al. 2001; Jenkins et al. 2007b; Sacks et al. 2001; Willcox et al. 2009). Healthy fat intake is very likely one mechanism for reducing CVD risk factors, however, other mechanisms, such as the high amounts of phytochemicals, high antioxidant intake, low glycemic load and resultant lowered oxidative stress are also likely playing a role in reducing risk for cardiovascular disease and other age-associated diseases. A comparison of the nutrient profiles of these dietary patterns in Table 1 showed that the traditional Okinawan diet is the lowest in fat, particularly in terms of saturated fat, and.A into oligomers and has a clearance effect on the existing A (Cole et al. 2007). A very interesting in vivo approach with multiphoton microscopy showed the ability of curcumin to cross the blood-brain barrier (BBB) and disrupt amyloid plaques (GarciaAlloza et al. 2007). Interestingly, curcumin possesses both MAO-A- and MAO-B-inhibiting properties and has been shown to modulate the levels of noradrenaline, dopamine and serotonin in the brain, demonstrating antidepressant effects in animal models of depression (Scapagnini et al. 2012) and in patients with major depressive disorder (Sanmukhani et al. 2013). Much of the research conducted to date on curcumin has been focused on exploring its protective and therapeutic effects against age-related degeneration. Recently the possibility that curcumin and its metabolites can modulate pathways directly involved in the determination of lifespan and extension of longevity, has been also highlighted (Shen et al. 2013). Tetrahydrocurcumin (THC), an active metabolite of curcumin, produced after its ingestion, has been shown to extend lifespan of drosophila under normal conditions, by attenuating oxidative stress via FOXO and Sir2 modulation (Xiang et al. 2011). Curcuminoids may also affect mammalian longevity, as shown in mice fed diets containing THC starting at the age of 13 months, which showed significantly increased mean lifespan (Shen et al. 2013).Author Manuscript Author Manuscript Author Manuscript Author ManuscriptSummary and ConclusionsThe traditional diet in Okinawa is based on green and yellow vegetables, root vegetables (principally sweet potatoes), soybean-based foods, and other plants, many with medicinal properties. This is supplemented by regular seafood consumption and consumption of smaller amounts of lean meats, fruit, and medicinal garnishes and spices. Sanpin (jasmine) tea is the principal beverage, consumed with meals and awamori (Okinawan sake) is the social drink of choice. The dietary composition over the past half-century has changed from a low calorie diet dominated by low glycemic index carbohydrates, low in protein and fat, to one more moderate in all three macronutrients. While the caloric content has increased due to higherMech Ageing Dev. Author manuscript; available in PMC 2017 April 24.Willcox et al.Pageconsumption of calorically dense foods, the diet remains very healthy by most expert criteria including the National Cholesterol Education Program (NCEP), the U.S. Dietary Guidelines for Americans Advisory Committee, and the Unified Dietary Guidelines. Many of the characteristics of the traditional Okinawan diet are shared with other healthy dietary patterns, such as the traditional Mediterranean diet, the modern DASH diet, and the modern Portfolio diet. All these dietary patterns have been found to be associated with reduced risk for cardiovascular disease (Appel, 2008; Fung et al. 2001; Jenkins et al. 2007b; Sacks et al. 2001; Willcox et al. 2009). Healthy fat intake is very likely one mechanism for reducing CVD risk factors, however, other mechanisms, such as the high amounts of phytochemicals, high antioxidant intake, low glycemic load and resultant lowered oxidative stress are also likely playing a role in reducing risk for cardiovascular disease and other age-associated diseases. A comparison of the nutrient profiles of these dietary patterns in Table 1 showed that the traditional Okinawan diet is the lowest in fat, particularly in terms of saturated fat, and.

Rtive treatment reduced anger, but DBT did not. Furthermore, only TFP

Rtive treatment reduced anger, but DBT did not. Furthermore, only TFP was associated with significant reductions in irritability, physical assault and verbal aggression. Findings indicate that all three treatments are effective in reducing symptoms and dysfunction associated with BPD. Consistent with previous findings, DBT did have a positive effect on suicide-related outcomes. However, the most BMS-791325 web widespread gains were observed among clients in TFP. In another study, McMain and colleagues (27) compared DBT (n = 90) to general psychiatric management (n = 90), which was based on the APA recommendations, and consisted of psychodynamic psychotherapy and symptom-targeted medication management. From the baseline assessment to the end of treatment, both groups showed significant improvements in almost every outcome assessedPsychiatr Clin North Am. Author manuscript; available in PMC 2011 September 1.NIH-PA Author Manuscript NIH-PA Author Manuscript NIH-PA Author ManuscriptMatusiewicz et al.Page(e.g., frequency of suicide attempts and non-suicidal self-injury, medical severity of these behaviors, emergency room visits and inpatient days, depression, anger, BPD symptom severity and overall symptom distress). However, contrary to predictions, the groups did not differ significantly on any treatment outcome, suggesting that DBT and general psychiatric management are equally effective in addressing symptoms and impairment associated with BPD. Taken together, findings from RCTs for DBT provide considerable support for its effectiveness as a treatment for BPD across many symptom domains. There is consistent evidence that DBT reduces suicidal parasuicidal behavior, decreases the medical risk associated with these behaviors, and produces fewer emergency visits and inpatient days. There is also evidence that DBT reduces affective symptoms of BPD (e.g., depression, anxiety, anger), and that it enhances global adjustment. It is also noteworthy that the effectiveness of DBT has been demonstrated in a range of real-world clinical settings, including a veteran’s affairs hospital (23), community mental health centers (28, 29), a university training clinic (30), and among clinicians in private practice (24, 26). Moreover, DBT has been found to be superior to treatment as usual, and generally equivalent to other active, structured, theoretically-sound outpatient treatments. Whereas standard DBT was developed to be a long-term outpatient treatment, there have been efforts to adapt DBT for use inpatients with BPD. In an initial trial, Barley and colleagues (31) compared frequency of non-suicidal self-injury and overdose before and after a long-term inpatient ward transitioned to DBT. As an additional control, they compared these changes to another general psychotherapy ward. They reported significant reductions in the incidence of non-suicidal self-injury, and parasuicidal behavior decreased on the DBT unit, whereas no decrease was observed on the comparison unit. Bohus and colleagues (32, 33) found similarly promising GGTI298 web outcomes following three-month inpatient DBT-based treatment, designed to jumpstart outpatient DBT. Inpatient DBT consisted of psychoeducation about BPD and mechanisms of treatment, skills training, and contingency management for parasuicidal behavior. In a pilot study, 24 female inpatients were assessed before and after 12 weeks of treatment. Significant improvements were observed in frequency of parasuicidal behavior, depression, anxiety, stress an.Rtive treatment reduced anger, but DBT did not. Furthermore, only TFP was associated with significant reductions in irritability, physical assault and verbal aggression. Findings indicate that all three treatments are effective in reducing symptoms and dysfunction associated with BPD. Consistent with previous findings, DBT did have a positive effect on suicide-related outcomes. However, the most widespread gains were observed among clients in TFP. In another study, McMain and colleagues (27) compared DBT (n = 90) to general psychiatric management (n = 90), which was based on the APA recommendations, and consisted of psychodynamic psychotherapy and symptom-targeted medication management. From the baseline assessment to the end of treatment, both groups showed significant improvements in almost every outcome assessedPsychiatr Clin North Am. Author manuscript; available in PMC 2011 September 1.NIH-PA Author Manuscript NIH-PA Author Manuscript NIH-PA Author ManuscriptMatusiewicz et al.Page(e.g., frequency of suicide attempts and non-suicidal self-injury, medical severity of these behaviors, emergency room visits and inpatient days, depression, anger, BPD symptom severity and overall symptom distress). However, contrary to predictions, the groups did not differ significantly on any treatment outcome, suggesting that DBT and general psychiatric management are equally effective in addressing symptoms and impairment associated with BPD. Taken together, findings from RCTs for DBT provide considerable support for its effectiveness as a treatment for BPD across many symptom domains. There is consistent evidence that DBT reduces suicidal parasuicidal behavior, decreases the medical risk associated with these behaviors, and produces fewer emergency visits and inpatient days. There is also evidence that DBT reduces affective symptoms of BPD (e.g., depression, anxiety, anger), and that it enhances global adjustment. It is also noteworthy that the effectiveness of DBT has been demonstrated in a range of real-world clinical settings, including a veteran’s affairs hospital (23), community mental health centers (28, 29), a university training clinic (30), and among clinicians in private practice (24, 26). Moreover, DBT has been found to be superior to treatment as usual, and generally equivalent to other active, structured, theoretically-sound outpatient treatments. Whereas standard DBT was developed to be a long-term outpatient treatment, there have been efforts to adapt DBT for use inpatients with BPD. In an initial trial, Barley and colleagues (31) compared frequency of non-suicidal self-injury and overdose before and after a long-term inpatient ward transitioned to DBT. As an additional control, they compared these changes to another general psychotherapy ward. They reported significant reductions in the incidence of non-suicidal self-injury, and parasuicidal behavior decreased on the DBT unit, whereas no decrease was observed on the comparison unit. Bohus and colleagues (32, 33) found similarly promising outcomes following three-month inpatient DBT-based treatment, designed to jumpstart outpatient DBT. Inpatient DBT consisted of psychoeducation about BPD and mechanisms of treatment, skills training, and contingency management for parasuicidal behavior. In a pilot study, 24 female inpatients were assessed before and after 12 weeks of treatment. Significant improvements were observed in frequency of parasuicidal behavior, depression, anxiety, stress an.

Ting both striated surfaces (Fig. 88 g); fore wing length almost always

Ting both striated surfaces (Fig. 88 g); fore wing length almost always 5.0 mm or more (range: 4.8?.1 mm); body length 4.5 mm (range: 4.1?.9 mm) [Hosts: Quadrus cerialis. A total of 22 diagnostic characters in the barcoding region: 67 C, 124 C, 133 T, 139 T, 181 A, 194 C, 200 T, 278 T, 298 A, 300 A, 311 G, 319 A, 335 A, 340 T, 346 T, 347 T, 523 C, 595 T, 616 T, 628 A, 634 T, 640 C] . ………………………………….Apanteles manuelriosi Fern dez-Triana, sp. n.?2(1)?Review of Apanteles sensu stricto (Hymenoptera, Braconidae, Microgastrinae)…carlosguadamuzi species-group This group comprises six species with extensive LLY-507MedChemExpress LLY-507 yellow-orange coloration, smooth mesoscutellar disc, mediotergite 1 weakly sculptured and light coloured with orangeyellow to light brown (males tend to have tergites with darker coloration, compared to females). The group is strongly supported by the Bayesian molecular analysis (PP: 1.0, Fig. 1). Hosts: mostly Crambidae, but some species reared from Choreutidae, Elachistidae, and Gelechiidae. Some species are Serabelisib dose gregarious and some are solitary parasitoids. All described species are from ACG, although we have seen undescribed species from other Neotropical areas. Key to species of the carlosguadamuzi group 1 ?2(1) ?3(1) ?4(3) ?5(3) T1 light brown, distinctly darker than T2 (Figs 91 g, 93 f) [Host: Ategumia lotanalis] ………………………………………………………………………………………..2 T1 entirely orange or orange-yellow, same color as T2 (Figs 90 g, 92 f, 94 f) …. 3 Fore wing with vein r 1.8?.0 ?as long as vein 2RS, and vein 2RS 1.0 ?as long as vein 2M ….Apanteles cinthiabarrantesae Fern dez-Triana, sp. n. Fore wing with vein r 1.3 ?as long as vein 2RS, and vein 2RS 1.6 ?as long as vein 2M ……………..Apanteles javiercontrerasi Fern dez-Triana, sp. n. T2 width at posterior margin at most 3.1 ?its median length (Fig. 94 f); ocular-ocellar line at most 1.8 ?posterior ocellus diameter …………………….4 T2 width at posterior margin at least 3.9 ?its median length (Figs 90 g, 92 f); ocular-ocellar line at least 2.1 ?posterior ocellus diameter …………………5 T1 2.5 ?as long as wide at posterior margin; T2 width at posterior margin 3.1 ?median length; fore wing with vein 2RS 1.6 ?as long as vein 2M [Hosts: Gelechiidae] …………..Apanteles jesusbrenesi Fern dez-Triana, sp. n. (N=4) T1 3.1 ?as long as wide at posterior margin; T2 width at posterior margin 2.7 ?median length; fore wing with vein 2RS 1.9 ?as long as vein 2M [Hosts: Elachistidae] ……Apanteles williamcamposi Fern dez-Triana, sp. n. (N=2) Metatarsus, posterior 0.3 of metatibia, and posterior 0.1 of metafemur brown to black, contrasting with rest of hind leg which is orange-yellow; body length 3.2?.4 mm; fore wing length 3.4?.6 mm; fore wing with vein r 2.1 ?as long as 2RS; flagellomerus 2 2.6 ?as long as wide; metafemur 3.2 ?as long as wide [Hosts: Choreutidae, Crambidae] …………………………………………….. …………………Apanteles carlosguadamuzi Fern dez-Triana, sp. n. (N=5) Metatarsus yellow or orange-yellow, same color as rest of hind leg, except for 0.2 or less of metatibia which is brown; body length usually 2.5?.7 mm (rarely up to 3.0 mm); fore wing length 2.7?.9 mm (rarely up to 3.2 mm); fore wing with vein r 1.3 ?as long as 2RS; flagellomerus 2 3.2 ?as long as wide; metafemur 2.9 ?as long as wide [Hosts: Crambidae] …………………….. ……………………Ting both striated surfaces (Fig. 88 g); fore wing length almost always 5.0 mm or more (range: 4.8?.1 mm); body length 4.5 mm (range: 4.1?.9 mm) [Hosts: Quadrus cerialis. A total of 22 diagnostic characters in the barcoding region: 67 C, 124 C, 133 T, 139 T, 181 A, 194 C, 200 T, 278 T, 298 A, 300 A, 311 G, 319 A, 335 A, 340 T, 346 T, 347 T, 523 C, 595 T, 616 T, 628 A, 634 T, 640 C] . ………………………………….Apanteles manuelriosi Fern dez-Triana, sp. n.?2(1)?Review of Apanteles sensu stricto (Hymenoptera, Braconidae, Microgastrinae)…carlosguadamuzi species-group This group comprises six species with extensive yellow-orange coloration, smooth mesoscutellar disc, mediotergite 1 weakly sculptured and light coloured with orangeyellow to light brown (males tend to have tergites with darker coloration, compared to females). The group is strongly supported by the Bayesian molecular analysis (PP: 1.0, Fig. 1). Hosts: mostly Crambidae, but some species reared from Choreutidae, Elachistidae, and Gelechiidae. Some species are gregarious and some are solitary parasitoids. All described species are from ACG, although we have seen undescribed species from other Neotropical areas. Key to species of the carlosguadamuzi group 1 ?2(1) ?3(1) ?4(3) ?5(3) T1 light brown, distinctly darker than T2 (Figs 91 g, 93 f) [Host: Ategumia lotanalis] ………………………………………………………………………………………..2 T1 entirely orange or orange-yellow, same color as T2 (Figs 90 g, 92 f, 94 f) …. 3 Fore wing with vein r 1.8?.0 ?as long as vein 2RS, and vein 2RS 1.0 ?as long as vein 2M ….Apanteles cinthiabarrantesae Fern dez-Triana, sp. n. Fore wing with vein r 1.3 ?as long as vein 2RS, and vein 2RS 1.6 ?as long as vein 2M ……………..Apanteles javiercontrerasi Fern dez-Triana, sp. n. T2 width at posterior margin at most 3.1 ?its median length (Fig. 94 f); ocular-ocellar line at most 1.8 ?posterior ocellus diameter …………………….4 T2 width at posterior margin at least 3.9 ?its median length (Figs 90 g, 92 f); ocular-ocellar line at least 2.1 ?posterior ocellus diameter …………………5 T1 2.5 ?as long as wide at posterior margin; T2 width at posterior margin 3.1 ?median length; fore wing with vein 2RS 1.6 ?as long as vein 2M [Hosts: Gelechiidae] …………..Apanteles jesusbrenesi Fern dez-Triana, sp. n. (N=4) T1 3.1 ?as long as wide at posterior margin; T2 width at posterior margin 2.7 ?median length; fore wing with vein 2RS 1.9 ?as long as vein 2M [Hosts: Elachistidae] ……Apanteles williamcamposi Fern dez-Triana, sp. n. (N=2) Metatarsus, posterior 0.3 of metatibia, and posterior 0.1 of metafemur brown to black, contrasting with rest of hind leg which is orange-yellow; body length 3.2?.4 mm; fore wing length 3.4?.6 mm; fore wing with vein r 2.1 ?as long as 2RS; flagellomerus 2 2.6 ?as long as wide; metafemur 3.2 ?as long as wide [Hosts: Choreutidae, Crambidae] …………………………………………….. …………………Apanteles carlosguadamuzi Fern dez-Triana, sp. n. (N=5) Metatarsus yellow or orange-yellow, same color as rest of hind leg, except for 0.2 or less of metatibia which is brown; body length usually 2.5?.7 mm (rarely up to 3.0 mm); fore wing length 2.7?.9 mm (rarely up to 3.2 mm); fore wing with vein r 1.3 ?as long as 2RS; flagellomerus 2 3.2 ?as long as wide; metafemur 2.9 ?as long as wide [Hosts: Crambidae] …………………….. ……………………

Obasanjo Apc

Ptor (EGFR), the vascular endothelial growth aspect receptor (VEGFR), or the platelet-derived development aspect receptor (PDGFR) loved ones. All receptor tyrosine kinases (RTK) are transmembrane proteins, whose amino-terminal end is extracellular (transmembrane proteins variety I). Their basic structure is comprised of an extracellular ligandbinding domain (ectodomain), a little hydrophobic transmembrane domain plus a cytoplasmic domain, which contains a conserved area with tyrosine kinase activity. This area consists of two lobules (N-terminal and C-terminal) that form a hinge where the ATP necessary for the catalytic reactions is located [10]. Activation of RTK requires place upon ligand binding at the extracellular level. This binding induces oligomerization of receptor monomers, typically dimerization. Within this phenomenon, juxtaposition of your tyrosine-kinase domains of each receptors stabilizes the kinase active state [11]. Upon kinase activation, every single monomer phosphorylates tyrosine residues in the cytoplasmic tail from the opposite monomer (trans-phosphorylation). Then, these phosphorylated residues are recognized by cytoplasmic proteins containing Src homology-2 (SH2) or phosphotyrosine-binding (PTB) domains, triggering different signaling cascades. Cytoplasmic proteins with SH2 or PTB domains might be effectors, proteins with enzymatic activity, or adaptors, proteins that mediate the activation of enzymes lacking these recognition web pages. Some examples of signaling molecules are: phosphoinositide 3-kinase (PI3K), phospholipase C (PLC), development issue receptor-binding protein (Grb), or the kinase Src, The principle signaling pathways activated by RTK are: PI3K/Akt, Ras/Raf/ERK1/2 and signal transduction and activator of transcription (STAT) pathways (Figure 1).Cells 2014, 3 Figure 1. Principal signal transduction pathways initiated by RTK.The PI3K/Akt pathway participates in apoptosis, migration and cell invasion manage [12]. This signaling cascade is initiated by PI3K activation because of RTK phosphorylation. PI3K phosphorylates phosphatidylinositol 4,5-bisphosphate (PIP2) creating phosphatidylinositol three,four,5-triphosphate (PIP3), which mediates the activation of your serine/threonine kinase Akt (also called protein kinase B). PIP3 induces Akt anchorage to the cytosolic side of PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/20502316/ the plasma membrane, where the phosphoinositide-dependent protein kinase 1 (PDK1) and also the phosphoinositide-dependent protein kinase 2 (PDK2) activate Akt by phosphorylating threonine 308 and serine 473 residues, respectively. The when elusive PDK2, on the other hand, has been recently identified as mammalian target of rapamycin (mTOR) within a rapamycin-insensitive complicated with rictor and Sin1 [13]. Upon phosphorylation, Akt is able to phosphorylate a plethora of substrates involved in cell cycle regulation, apoptosis, protein synthesis, glucose metabolism, and so forth [12,14]. A frequent order A-196 alteration located in glioblastoma that impacts this signaling pathway is mutation or genetic loss of your tumor suppressor gene PTEN (Phosphatase and Tensin homologue deleted on chromosome ten), which encodes a dual-specificity protein phosphatase that catalyzes PIP3 dephosphorylation [15]. As a result, PTEN is actually a essential negative regulator from the PI3K/Akt pathway. About 20 to 40 of glioblastomas present PTEN mutational inactivation [16] and about 35 of glioblastomas endure genetic loss because of promoter methylation [17]. The Ras/Raf/ERK1/2 pathway is definitely the primary mitogenic route initiated by RTK. This signaling pathway is trig.

Phosphorylation Of Crtc3 By The Salt-Inducible Kinases

Ptor (EGFR), the vascular endothelial development aspect receptor (VEGFR), or the platelet-derived development factor receptor (PDGFR) family. All receptor tyrosine kinases (RTK) are transmembrane proteins, whose amino-terminal end is extracellular (transmembrane proteins type I). Their common structure is comprised of an extracellular ligandbinding domain (ectodomain), a little hydrophobic transmembrane domain as well as a cytoplasmic domain, which consists of a conserved region with tyrosine kinase activity. This area consists of two lobules (N-terminal and C-terminal) that kind a hinge where the ATP required for the catalytic reactions is situated [10]. Activation of RTK requires place upon ligand binding in the extracellular level. This binding induces oligomerization of receptor monomers, normally dimerization. Within this phenomenon, juxtaposition from the tyrosine-kinase domains of both receptors stabilizes the kinase active state [11]. Upon kinase activation, each and every monomer phosphorylates tyrosine residues inside the cytoplasmic tail on the opposite monomer (trans-phosphorylation). Then, these phosphorylated residues are recognized by cytoplasmic proteins containing Src homology-2 (SH2) or phosphotyrosine-binding (PTB) domains, triggering various signaling cascades. Cytoplasmic proteins with SH2 or PTB domains can be effectors, proteins with enzymatic activity, or adaptors, proteins that mediate the activation of enzymes lacking these recognition sites. Some examples of signaling molecules are: phosphoinositide 3-kinase (PI3K), phospholipase C (PLC), growth factor receptor-binding protein (Grb), or the kinase Src, The principle signaling pathways activated by RTK are: PI3K/Akt, Ras/Raf/ERK1/2 and signal transduction and activator of transcription (STAT) pathways (Figure 1).Cells 2014, 3 Figure 1. Primary signal transduction pathways initiated by RTK.The PI3K/Akt pathway participates in apoptosis, migration and cell invasion manage [12]. This signaling cascade is initiated by PI3K activation due to RTK phosphorylation. PI3K phosphorylates phosphatidylinositol 4,5-bisphosphate (PIP2) creating phosphatidylinositol 3,4,5-triphosphate (PIP3), which mediates the activation of the serine/threonine kinase Akt (also referred to as protein kinase B). PIP3 induces Akt anchorage to the cytosolic side of PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/20502316/ the plasma membrane, buy Go 6850 exactly where the phosphoinositide-dependent protein kinase 1 (PDK1) along with the phosphoinositide-dependent protein kinase 2 (PDK2) activate Akt by phosphorylating threonine 308 and serine 473 residues, respectively. The once elusive PDK2, nonetheless, has been recently identified as mammalian target of rapamycin (mTOR) in a rapamycin-insensitive complex with rictor and Sin1 [13]. Upon phosphorylation, Akt is in a position to phosphorylate a plethora of substrates involved in cell cycle regulation, apoptosis, protein synthesis, glucose metabolism, and so forth [12,14]. A frequent alteration found in glioblastoma that affects this signaling pathway is mutation or genetic loss of your tumor suppressor gene PTEN (Phosphatase and Tensin homologue deleted on chromosome ten), which encodes a dual-specificity protein phosphatase that catalyzes PIP3 dephosphorylation [15]. Hence, PTEN is actually a crucial adverse regulator in the PI3K/Akt pathway. About 20 to 40 of glioblastomas present PTEN mutational inactivation [16] and about 35 of glioblastomas endure genetic loss as a result of promoter methylation [17]. The Ras/Raf/ERK1/2 pathway is definitely the primary mitogenic route initiated by RTK. This signaling pathway is trig.

Zelda Apc

Ptor (EGFR), the vascular endothelial growth aspect receptor (VEGFR), or the platelet-derived growth element receptor (PDGFR) family. All receptor tyrosine kinases (RTK) are transmembrane proteins, whose amino-terminal finish is extracellular (transmembrane proteins sort I). Their common structure is comprised of an extracellular ligandbinding domain (ectodomain), a little hydrophobic transmembrane domain as well as a cytoplasmic domain, which includes a conserved area with tyrosine kinase activity. This area consists of two lobules (N-terminal and C-terminal) that form a hinge where the ATP needed for the catalytic reactions is located [10]. order Podocarpusflavone A activation of RTK takes spot upon ligand binding at the extracellular level. This binding induces oligomerization of receptor monomers, generally dimerization. Within this phenomenon, juxtaposition of the tyrosine-kinase domains of both receptors stabilizes the kinase active state [11]. Upon kinase activation, every monomer phosphorylates tyrosine residues in the cytoplasmic tail from the opposite monomer (trans-phosphorylation). Then, these phosphorylated residues are recognized by cytoplasmic proteins containing Src homology-2 (SH2) or phosphotyrosine-binding (PTB) domains, triggering different signaling cascades. Cytoplasmic proteins with SH2 or PTB domains might be effectors, proteins with enzymatic activity, or adaptors, proteins that mediate the activation of enzymes lacking these recognition sites. Some examples of signaling molecules are: phosphoinositide 3-kinase (PI3K), phospholipase C (PLC), development element receptor-binding protein (Grb), or the kinase Src, The primary signaling pathways activated by RTK are: PI3K/Akt, Ras/Raf/ERK1/2 and signal transduction and activator of transcription (STAT) pathways (Figure 1).Cells 2014, 3 Figure 1. Principal signal transduction pathways initiated by RTK.The PI3K/Akt pathway participates in apoptosis, migration and cell invasion control [12]. This signaling cascade is initiated by PI3K activation due to RTK phosphorylation. PI3K phosphorylates phosphatidylinositol 4,5-bisphosphate (PIP2) generating phosphatidylinositol 3,4,5-triphosphate (PIP3), which mediates the activation of the serine/threonine kinase Akt (also known as protein kinase B). PIP3 induces Akt anchorage towards the cytosolic side of PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/20502316/ the plasma membrane, exactly where the phosphoinositide-dependent protein kinase 1 (PDK1) plus the phosphoinositide-dependent protein kinase 2 (PDK2) activate Akt by phosphorylating threonine 308 and serine 473 residues, respectively. The after elusive PDK2, nonetheless, has been not too long ago identified as mammalian target of rapamycin (mTOR) in a rapamycin-insensitive complex with rictor and Sin1 [13]. Upon phosphorylation, Akt is able to phosphorylate a plethora of substrates involved in cell cycle regulation, apoptosis, protein synthesis, glucose metabolism, and so forth [12,14]. A frequent alteration found in glioblastoma that affects this signaling pathway is mutation or genetic loss in the tumor suppressor gene PTEN (Phosphatase and Tensin homologue deleted on chromosome ten), which encodes a dual-specificity protein phosphatase that catalyzes PIP3 dephosphorylation [15]. As a result, PTEN is often a crucial damaging regulator of your PI3K/Akt pathway. About 20 to 40 of glioblastomas present PTEN mutational inactivation [16] and about 35 of glioblastomas endure genetic loss as a consequence of promoter methylation [17]. The Ras/Raf/ERK1/2 pathway will be the principal mitogenic route initiated by RTK. This signaling pathway is trig.

Phosphorylation Of Crtc3 By The Salt-Inducible Kinases

Ptor (EGFR), the vascular endothelial development aspect receptor (VEGFR), or the platelet-derived development issue receptor (PDGFR) family. All receptor tyrosine kinases (RTK) are transmembrane proteins, whose amino-terminal finish is extracellular (transmembrane proteins type I). Their general structure is comprised of an extracellular ligandbinding domain (ectodomain), a small hydrophobic transmembrane domain as well as a cytoplasmic domain, which consists of a conserved area with tyrosine kinase activity. This region consists of two lobules (N-terminal and C-terminal) that form a hinge where the ATP required for the catalytic reactions is situated [10]. Activation of RTK requires place upon ligand binding in the extracellular level. This binding induces oligomerization of receptor monomers, usually dimerization. In this phenomenon, juxtaposition from the tyrosine-kinase domains of both receptors stabilizes the kinase active state [11]. Upon kinase activation, each and every monomer phosphorylates tyrosine residues inside the cytoplasmic tail of your opposite monomer (trans-phosphorylation). Then, these phosphorylated residues are recognized by cytoplasmic proteins containing Src homology-2 (SH2) or phosphotyrosine-binding (PTB) domains, triggering distinctive signaling cascades. Cytoplasmic proteins with SH2 or PTB domains is usually effectors, proteins with enzymatic activity, or adaptors, proteins that mediate the activation of enzymes ARS-853 supplier lacking these recognition web-sites. Some examples of signaling molecules are: phosphoinositide 3-kinase (PI3K), phospholipase C (PLC), development element receptor-binding protein (Grb), or the kinase Src, The primary signaling pathways activated by RTK are: PI3K/Akt, Ras/Raf/ERK1/2 and signal transduction and activator of transcription (STAT) pathways (Figure 1).Cells 2014, three Figure 1. Principal signal transduction pathways initiated by RTK.The PI3K/Akt pathway participates in apoptosis, migration and cell invasion handle [12]. This signaling cascade is initiated by PI3K activation as a consequence of RTK phosphorylation. PI3K phosphorylates phosphatidylinositol four,5-bisphosphate (PIP2) making phosphatidylinositol 3,four,5-triphosphate (PIP3), which mediates the activation in the serine/threonine kinase Akt (also known as protein kinase B). PIP3 induces Akt anchorage for the cytosolic side of PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/20502316/ the plasma membrane, exactly where the phosphoinositide-dependent protein kinase 1 (PDK1) and the phosphoinositide-dependent protein kinase two (PDK2) activate Akt by phosphorylating threonine 308 and serine 473 residues, respectively. The after elusive PDK2, however, has been recently identified as mammalian target of rapamycin (mTOR) within a rapamycin-insensitive complicated with rictor and Sin1 [13]. Upon phosphorylation, Akt is capable to phosphorylate a plethora of substrates involved in cell cycle regulation, apoptosis, protein synthesis, glucose metabolism, and so forth [12,14]. A frequent alteration discovered in glioblastoma that impacts this signaling pathway is mutation or genetic loss in the tumor suppressor gene PTEN (Phosphatase and Tensin homologue deleted on chromosome ten), which encodes a dual-specificity protein phosphatase that catalyzes PIP3 dephosphorylation [15]. As a result, PTEN is usually a crucial adverse regulator on the PI3K/Akt pathway. About 20 to 40 of glioblastomas present PTEN mutational inactivation [16] and about 35 of glioblastomas endure genetic loss because of promoter methylation [17]. The Ras/Raf/ERK1/2 pathway could be the most important mitogenic route initiated by RTK. This signaling pathway is trig.

G then able to bind inner PM phospholipids as well as

G then able to bind inner PM phospholipids as well as cytoplasmic membranes of organelles (Fig. 3d; Table 1); and/or (ii) incubated with cells to target outer SC144 structure leaflet phospholipids after transbilayer flip-flop. The pleckstrin homology (PH) domain is one of these well-characterized probes specific for phosphoinositides (PIs; [122]). The 100 amino acid-PH domain is contained in several proteins, such as pleckstrin or phospholipase C (PLC), with distinct binding affinity for different PIs [123]. For instance, PH domain of PLC (PH-PLC) has a high affinity for phosphatidylinositol-4,5-bisphosphate (PIP2) [124, 125]. The discoidin C2 domain is another probe, specific for phosphatidylserine (PS). The 160 amino acid-discoidin C2 domain is present in blood coagulation factors V and VIII, milk fat globule-EGF factor 8 (MFGE8; also known as lactadherin [Lact-C2]) and other plasma proteins. PH or discoidin C2 domains can be fluorescently tagged, allowing to study phospholipid membrane distribution [126-128]. Other globular domains capable to bind phospholipids at the membrane surface include: (i) the FYVE zinc Pan-RAS-IN-1 clinical trials finger domain found in EEA1 (Early Endosome Antigen 1) a.o. that binds to phosphatidylinositol-3-phosphate (PI3P); and (ii) the calcium-dependent phospholipid binding Annexins, such as Annexin A2, which preferentially interacts with PIP2, or Annexin A5, which is currently the most commonly used probe for PS targeting at outer PM leaflet [129]. To further overcome limitation due to lack of PS labeling at the luminal membrane leaflet of organelles. Parton and coll. recently developed a novel on-section labeling approach on fast-frozen sample using purified GST (glutathione-S-transferase)-Lact-C2 fusion protein followed by transmission electron microscopy. This technique is based on high-pressure freezing, freeze-substitution with minimal fixatives and embedding at low temperature. Sections are then fixed, labeled with purified GST-Lact-C2 and followed by detection with anti-GST antibody and protein A?Author Manuscript Author Manuscript Author Manuscript Author ManuscriptProg Lipid Res. Author manuscript; available in PMC 2017 April 01.Carquin et al.Pagegold. Such method avoids cell permeabilization as well as detergent extraction [126]. For more details on phospholipid-binding domains, please refer to [130]. Similarly to other probes, this approach also presents limitations including perturbation of normal lipid function upon high expression and high variability of affinity and specificity [129, 131]. 3.1.3. Antibodies, Fab fragments and nanobodies–Antibodies have been recognized as gold standard to detect proteins. Interestingly, several antibodies have also been generated to decorate PM lipids (Fig. 3e). For example, there are monoclonal antibodies (mAbs) produced to detect specific GSLs expressed during the differentiation of oligodendrocytes and used for studying their in vitro maturation: (i) the mAb A2B5, against gangliosides GD3, GT3 and O-acetylated GT3 in early oligodendrocyte progenitors; (ii) the mAb O4, against sulfated GSLs expressed by late progenitors; and (iii) the mAb O1 and the mAb Ranscht, against galactosylceramides in mature oligodendrocytes (for a review, see [132]). These antibodies have revealed submicrometric GSL-enriched domains at different stages of oligodendrocyte differentiation, as illustrated in Table 1. Although less developed, antibodies are also used to decorate phospholipids. For example, the role of PS do.G then able to bind inner PM phospholipids as well as cytoplasmic membranes of organelles (Fig. 3d; Table 1); and/or (ii) incubated with cells to target outer leaflet phospholipids after transbilayer flip-flop. The pleckstrin homology (PH) domain is one of these well-characterized probes specific for phosphoinositides (PIs; [122]). The 100 amino acid-PH domain is contained in several proteins, such as pleckstrin or phospholipase C (PLC), with distinct binding affinity for different PIs [123]. For instance, PH domain of PLC (PH-PLC) has a high affinity for phosphatidylinositol-4,5-bisphosphate (PIP2) [124, 125]. The discoidin C2 domain is another probe, specific for phosphatidylserine (PS). The 160 amino acid-discoidin C2 domain is present in blood coagulation factors V and VIII, milk fat globule-EGF factor 8 (MFGE8; also known as lactadherin [Lact-C2]) and other plasma proteins. PH or discoidin C2 domains can be fluorescently tagged, allowing to study phospholipid membrane distribution [126-128]. Other globular domains capable to bind phospholipids at the membrane surface include: (i) the FYVE zinc finger domain found in EEA1 (Early Endosome Antigen 1) a.o. that binds to phosphatidylinositol-3-phosphate (PI3P); and (ii) the calcium-dependent phospholipid binding Annexins, such as Annexin A2, which preferentially interacts with PIP2, or Annexin A5, which is currently the most commonly used probe for PS targeting at outer PM leaflet [129]. To further overcome limitation due to lack of PS labeling at the luminal membrane leaflet of organelles. Parton and coll. recently developed a novel on-section labeling approach on fast-frozen sample using purified GST (glutathione-S-transferase)-Lact-C2 fusion protein followed by transmission electron microscopy. This technique is based on high-pressure freezing, freeze-substitution with minimal fixatives and embedding at low temperature. Sections are then fixed, labeled with purified GST-Lact-C2 and followed by detection with anti-GST antibody and protein A?Author Manuscript Author Manuscript Author Manuscript Author ManuscriptProg Lipid Res. Author manuscript; available in PMC 2017 April 01.Carquin et al.Pagegold. Such method avoids cell permeabilization as well as detergent extraction [126]. For more details on phospholipid-binding domains, please refer to [130]. Similarly to other probes, this approach also presents limitations including perturbation of normal lipid function upon high expression and high variability of affinity and specificity [129, 131]. 3.1.3. Antibodies, Fab fragments and nanobodies–Antibodies have been recognized as gold standard to detect proteins. Interestingly, several antibodies have also been generated to decorate PM lipids (Fig. 3e). For example, there are monoclonal antibodies (mAbs) produced to detect specific GSLs expressed during the differentiation of oligodendrocytes and used for studying their in vitro maturation: (i) the mAb A2B5, against gangliosides GD3, GT3 and O-acetylated GT3 in early oligodendrocyte progenitors; (ii) the mAb O4, against sulfated GSLs expressed by late progenitors; and (iii) the mAb O1 and the mAb Ranscht, against galactosylceramides in mature oligodendrocytes (for a review, see [132]). These antibodies have revealed submicrometric GSL-enriched domains at different stages of oligodendrocyte differentiation, as illustrated in Table 1. Although less developed, antibodies are also used to decorate phospholipids. For example, the role of PS do.