Brain-derived neurotrophic factor (BDNF) regulates dendritic branching and dendritic spine morphology, as well as synaptic plasticity and long-term potentiation. Consequently, BDNF deficiency has ...been associated with some neurological disorders such as Alzheimer's, Parkinson's or Huntington's diseases. In contrast, elevated BDNF levels correlate with recovery after traumatic central nervous system (CNS) injuries. The utility of BDNF as a therapeutic agent is limited by its short half-life in a pathological microenvironment and its low efficacy caused by unwanted consumption of non-neuronal cells or inappropriate dosing. Here, we tested the activity of chitosan microsphere-encapsulated BDNF to prevent clearance and prolong the efficacy of this neurotrophin. Neuritic growth activity of BDNF release from chitosan microspheres was observed in the PC12 rat pheochromocytoma cell line, which is dependent on neurotrophins to differentiate via the neurotrophin receptor (NTR). We obtained a rapid and sustained increase in neuritic out-growth of cells treated with BDNF-loaded chitosan microspheres over control cells (
< 0.001). The average of neuritic out-growth velocity was three times higher in the BDNF-loaded chitosan microspheres than in the free BDNF. We conclude that the slow release of BDNF from chitosan microspheres enhances signaling through NTR and promotes axonal growth in neurons, which could constitute an important therapeutic agent in neurodegenerative diseases and CNS lesions.
KRAS mutations in colorectal cancer primary tumors predict resistance to anti-Epidermal Growth Factor Receptor (EGFR) monoclonal antibody therapy in patients with metastatic colorectal cancer, and ...thus represent a true indicator of EGFR pathway activation status.
KRAS mutations were retrospectively studied using polymerase chain reactions and subsequent sequencing of codons 12 and 13 (exon 2) in 110 patients with metastatic colorectal tumors. These studies were performed using tissue samples from both the primary tumor and their related metastases (93 liver, 84%; 17 lung, 16%). All patients received adjuvant 5-Fluorouracil-based polychemotherapy after resection of metastases. None received anti-EGFR therapy. Mutations in KRAS were observed in 37 (34%) of primary tumors and in 40 (36%) of related metastases, yielding a 94% level of concordance (kappa index 0.86). Patients with primary tumors possessing KRAS mutations had a shorter disease-free survival period after metastasis resection (12.0 vs 18.0 months; P = 0.035) than those who did not. A higher percentage of KRAS mutations was detected in primary tumors of patiens with lung metastases than in patients with liver metastases (59% vs 32%; p = 0.054). To further evaluate this finding we analyzed 120 additional patients with unresectable metastatic colorectal cancer who previously had their primary tumors evaluated for KRAS mutational status for clinical purposes. Separately, the analysis of these 120 patients showed a tendency towards a higher degree of KRAS mutations in primary tumors of patients with lung metastases, although it did not reach statistical significance. Taken together the group of 230 patients showed that KRAS was mutated significantly more often in the primary tumors of patients with lung metastases (57% vs 35%; P = 0.006).
Our results suggest a role for KRAS mutations in the propensity of primary colorectal tumors to metastasize to the lung.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
The study of cross-reactivity in allergy is key to both understanding. the allergic response of many patients and providing them with a rational treatment In the present study, protein microarrays ...and a co-sensitization graph approach were used in conjunction with an allergen microarray immunoassay. This enabled us to include a wide number of proteins and a large number of patients, and to study sensitization profiles among members of the LTP family. Fourteen LTPs from the most frequent plant food-induced allergies in the geographical area studied were printed into a microarray specifically designed for this research. 212 patients with fruit allergy and 117 food-tolerant pollen allergic subjects were recruited from seven regions of Spain with different pollen profiles, and their sera were tested with allergen microarray. This approach has proven itself to be a good tool to study cross-reactivity between members of LTP family, and could become a useful strategy to analyze other families of allergens.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
To the Editor: Occupational asthma accounts for more than 10% of all cases of asthma in adults, and baker's asthma (BA) is the leading cause of occupational respiratory disease in Western countries.1 ...Such occupational allergic respiratory disorders are often misdiagnosed, with significant legal, economic, and health impacts for affected patients.1,2 The incidence of BA among young bakers has been reported to range from 0.3 to 2.4 cases per 1000 person-year, and an increasing number of asthma cases are being reported among supermarket bakery workers.3 In recent years, some studies aimed to determine the panel of wheat allergens for diagnosing patients with BA to avoid specific bronchial challenge with wheat due to its potential risk and technical requirements.4,5 Regarding in vitro diagnosis (specific IgE assays), all known diagnostic approaches have poor predictability and specificity.1,2 The introduction of microarray techniques featuring a large panel of purified allergens has been a major advance in the diagnosis of allergic diseases.6,7 However, this technique has been hardly applied to the diagnosis of patients with occupational asthma caused by wheat.7,8 In the present study, the allergen profiles of patients with BA from 3 different regions in Spain (Madrid, Malaga, and Valladolid) with a relevant bakery industry were characterized. Location Median age (y) (range) Sex (M/F) SPT wheat extract (positive) Bronchial challenge test (positive) Aeroallergen sensitization (SPT) Food allergy (SPT) Madrid n = 17 39.2 (28-64) 16/1 17/17 17/17 + Any SPT 88%Olive 35%Grass 29%Dust mites 35%Alternaria 11.7%Alpha-amylase 17.6% 53% No47% Yes18% Nuts6% Barley6% Egg6% Kiwi Malaga n = 10 38 (19-60) 8/2 10/10 10/10 + Any SPT 75%Olive 37.5%Grass 37.5%Dust mites 62.5%Alternaria 12.5%Alpha-amylase 12% 60% No40% Yes50% Peach25% Nuts25% Spices25% Barley Valladolid n = 18 31 (19-66) 14/4 18/18 18/18 + Any SPT 89%Olive 50%Grass 70%Dust mites 20%Alternaria 10%Alpha-amylase 22.4% 62% No38% Yes50% Barley25% Apple25% Peach Table I Clinical data of patients with BA included in the study F, Female; M, male.
Background
Obesity-associated dysfunctional intestinal permeability contributes to systemic chronic inflammation leading to the development of metabolic diseases. The inflammasomes constitute ...essential components in the regulation of intestinal homeostasis. We aimed to determine the impact of the inflammasomes in the regulation of gut barrier dysfunction and metabolic inflammation in the context of obesity and type 2 diabetes (T2D).
Methods
Blood samples obtained from 80 volunteers (
n
= 20 normal weight,
n
= 21 OB without T2D,
n
= 39 OB with T2D) and a subgroup of jejunum samples were used in a case–control study. Circulating levels of intestinal damage markers and expression levels of inflammasomes as well as their main effectors (IL-1β and IL-18) and key inflammation-related genes were analyzed. The impact of inflammation-related factors, different metabolites and
Akkermansia muciniphila
in the regulation of inflammasomes and intestinal integrity genes was evaluated. The effect of blocking
NLRP6
by using siRNA in inflammation was also studied.
Results
Increased circulating levels (
P
< 0.01) of the intestinal damage markers endotoxin, LBP, and zonulin in patients with obesity decreased (
P
< 0.05) after weight loss. Patients with obesity and T2D exhibited decreased (
P
< 0.05) jejunum gene expression levels of
NLRP6
and its main effector
IL18
together with increased (
P
< 0.05) mRNA levels of inflammatory markers. We further showed that while NLRP6 was primarily localized in goblet cells, NLRP3 was localized in the intestinal epithelial cells. Additionally, decreased (
P
< 0.05) mRNA levels of
Nlrp1
,
Nlrp3
and
Nlrp6
in the small intestinal tract obtained from rats with diet-induced obesity were found.
NLRP6
expression was regulated by taurine, parthenolide and
A. muciniphila
in the human enterocyte cell line CCL-241. Finally, a significant decrease (
P
< 0.01) in the expression and release of
MUC2
after the knockdown of
NLRP6
was observed.
Conclusions
The increased levels of intestinal damage markers together with the downregulation of
NLRP6
and
IL18
in the jejunum in obesity-associated T2D suggest a defective inflammasome sensing, driving to an impaired epithelial intestinal barrier that may regulate the progression of multiple obesity-associated comorbidities.
Cross-reactivity of plant foods is an important phenomenon in allergy, with geographical variations with respect to the number and prevalence of the allergens involved in this process, whose ...complexity requires detailed studies. We have addressed the role of thaumatin-like proteins (TLPs) in cross-reactivity between fruit and pollen allergies. A representative panel of 16 purified TLPs was printed onto an allergen microarray. The proteins selected belonged to the sources most frequently associated with peach allergy in representative regions of Spain. Sera from two groups of well characterized patients, one with allergy to Rosaceae fruit (FAG) and another against pollens but tolerant to food-plant allergens (PAG), were obtained from seven geographical areas with different environmental pollen profiles. Cross-reactivity between members of this family was demonstrated by inhibition assays. Only 6 out of 16 purified TLPs showed noticeable allergenic activity in the studied populations. Pru p 2.0201, the peach TLP (41%), chestnut TLP (24%) and plane pollen TLP (22%) proved to be allergens of probable relevance to fruit allergy, being mainly associated with pollen sensitization, and strongly linked to specific geographical areas such as Barcelona, Bilbao, the Canary Islands and Madrid. The patients exhibited >50% positive response to Pru p 2.0201 and to chestnut TLP in these specific areas. Therefore, their recognition patterns were associated with the geographical area, suggesting a role for pollen in the sensitization of these allergens. Finally, the co-sensitizations of patients considering pairs of TLP allergens were analyzed by using the co-sensitization graph associated with an allergen microarray immunoassay. Our data indicate that TLPs are significant allergens in plant food allergy and should be considered when diagnosing and treating pollen-food allergy.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
In this paper, a green analytical methodology based on fluorescence derivatization is proposed for the anti-helminthic drug monitoring ivermectin as environmental emergent contaminant. After sample ...clean-up, ivermectin was converted into a highly fluorescent derivative through a catalytic oxidation process followed by dehydration and tautomerization. Under optimal experimental conditions, a linear response was obtained for ivermectin within the range 0.38–600 μg L−1, with detection and quantification limits of 0.11 and 0.38 μg L−1, both values are lower than other previously reported. This method has been applied for ivermectin determination in environmental water samples at trace levels, showing its potential for contamination monitoring.
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