Phytohemagglutinin from Phaseolus vulgaris enhances the lung cancer cell chemotherapy sensitivity by changing cell membrane permeability.

Chemotherapy is still a prevalent strategy for clinical lung cancer treatment. However, the inevitable emerged drug resistance has become a great hurdle to therapeutic effect. Studies have demonstrated that the primary cause of drug resistance is a decrease in the chemotherapeutic medicine concentration. Several lectins have been confirmed to be effective as chemotherapy adjuvants, enhancing the anti-tumor effects of chemotherapy drugs. Here, we combined phytohemagglutinin (PHA), which has been reported possess anti-tumor effects, with chemotherapy drugs Cisplatin (DDP) and Adriamycin (ADM) on lung cancer cells to detect the sensitivities of PHA as a chemotherapy adjuvant. Our results demonstrated that the PHA significantly enhanced the sensitivity of lung cancer cells to DDP and ADM, and Western blot showed that PHA combined with DDP or ADM enhance cytotoxic effects by inhibiting autophagy and promoting apoptosis. More importantly, we found PHA enhanced the chemotherapeutic drugs cytotoxicity by changing the cell membrane to increase the intracellular chemotherapeutic drugs concentration. Besides, the combination of PHA and ADM increased the ADM concentration in the multidrug-resistant strain A549-R cells and achieved the drug sensitization effect. Our results suggest that PHA combined with chemotherapy can be applied in the treatment of lung cancer cells and lung cancer multidrug-resistant strains, and provide a novel strategy for clinical tumor chemotherapy and a new idea to solve the problem of drug resistance in clinical lung cancer.

Projections from the five divisions of the orbital cortex to the thalamus in the rat.

The orbital cortex (ORB) of the rat consists of five divisions: the medial (MO), ventral (VO), ventrolateral (VLO), lateral (LO), and dorsolateral (DLO) orbital cortices. No previous report has comprehensively examined and compared projections from each division of the ORB to the thalamus. Using the anterograde anatomical tracer, Phaseolus vulgaris leucoagglutinin, we describe the efferent projections from the five divisions of the ORB to the thalamus in the rat. We demonstrated that, with some overlap, each division of the ORB distributed in a distinct (and unique) manner to nuclei of the thalamus. Overall, ORB projected to a relatively restricted number of sites in the thalamus, and strikingly distributed entirely to structures of the medial/midline thalamus, while completely avoiding lateral regions or principal nuclei of the thalamus. The main termination sites in the thalamus were the paratenial nucleus (PT) and nucleus reuniens (RE) of the midline thalamus, the medial (MDm) and central (MDc) divisions of the mediodorsal nucleus, the intermediodorsal nucleus, the central lateral, paracentral, and central medial nuclei of the rostral intralaminar complex and the submedial nucleus (SM). With some exceptions, medial divisions of the ORB (MO, VO) mainly targeted "limbic-associated" nuclei such as PT, RE, and MDm, whereas lateral division (VLO, LO, DLO) primarily distributed to "sensorimotor-associated" nuclei including MDc, SM, and the rostral intralaminar complex. As discussed herein, the medial/midline thalamus may represent an important link (or bridge) between the orbital cortex and the hippocampus and between the ORB and medial prefrontal cortex. In summary, the present results demonstrate that each division of the orbital cortex projects in a distinct manner to nuclei of the thalamus which suggests unique functions for each division of the orbital cortex.

Characterization of the Transient Deficiency of PKC Isozyme Levels in Immature Cord Blood T Cells and Its Connection to Anti-Allergic Cytokine Profiles of the Matured Cells.

Cord blood T cells (CBTC) from a proportion of newborns express low/deficient levels of some protein kinase C (PKC) isozymes, with low levels of PKCζ correlating with increased risk of developing allergy and associated decrease in interferon-gamma (IFN-γ) producing T cells. Interestingly, these lower levels of PKCζ were increased/normalized by supplementing women during pregnancy with n-3 polyunsaturated fatty acids. However, at present, we have little understanding of the transient nature of the deficiency in the neonate and how PKCζ relates to other PKC isozymes and whether their levels influence maturation into IFN-γ producing T cells. There is also no information on PKCζ isozyme levels in the T cell subpopulations, CD4+ and CD8+ cells. These issues were addressed in the present study using a classical culture model of neonatal T cell maturation, initiated with phytohaemagglutinin (PHA) and recombinant human interleukin-2 (rhIL-2). Of the isozymes evaluated, PKCζ, ß2, δ, µ, ε, θ and λ/ι were low in CBTCs. The PKC isozyme deficiencies were also found in the CD4+ and CD8+ T cell subset levels of the PKC isozymes correlated between the two subpopulations. Examination of changes in the PKC isozymes in these deficient cells following addition of maturation signals showed a significant increase in expression within the first few hours for PKCζ, ß2 and µ, and 1-2 days for PKCδ, ε, θ and λ/ι. Only CBTC PKCζ isozyme levels correlated with cytokine production, with a positive correlation with IFN-γ, interleukin (IL)-2 and tumour necrosis factor-alpha (TNF), and a negative association with IL-9 and IL-10. The findings reinforce the specificity in using CBTC PKCζ levels as a biomarker for risk of allergy development and identify a period in which this can be potentially 'corrected' after birth.

Role of Phytomolecules in the Treatment of Obesity: Targets, Mechanisms and Limitations.

Obesity has become a worldwide health problem. It triggers additional co-morbidities like cardiovascular diseases, cancer, depression, sleep disorders, gastrointestinal problems and many more. Excess accumulation of fat in obesity could be caused by many factors like sedentary lifestyle, consumption of high-fat diet, genetic predisposition, etc. Imbalanced energy metabolism i.e., greater energy consumption than utilisation, invariably underlies obesity. Considering the high prevalence and continuous, uncontrolled increase of this major public health issue, there is an urgent need to find appropriate therapeutic agents with minimal or no side effects. The high prevalence of obesity in recent years has led to a surge in the number of drugs available in the market that claim to control obesity. Although there is a long list of medicines and management strategies that are available, selecting the right therapeutic intervention and feasible management of obesity is a challenge. Several phytochemicals like hydroxycitric acid, flavonoids, tannins, anthocyanins, phytohaemagglutinin, thymoquinone and epigallocatechin gallate have been shown to possess promising anti-obesity properties. However, studies providing information on how various phytochemicals exert their anti-obesity effects are inadequate. This calls for more experimentation in this less explored area of research. Additionally, the complication of obesity arises when it is a result of multiple factors and associated with a number of co-morbidities. In order to handle such complexities, combinatorial therapeutic interventions become effective. In this review, we have described the medicinal chemistry of different highly effective phytochemicals which can be used in the effective treatment and management of obesity.

Projections from the dorsomedial division of the bed nucleus of the stria terminalis to hypothalamic nuclei in the mouse.

As stressful environment is a potent modulator of feeding, we seek in the present work to decipher the neuroanatomical basis for an interplay between stress and feeding behaviors. For this, we combined anterograde and retrograde tracing with immunohistochemical approaches to investigate the patterns of projections between the dorsomedial division of the bed nucleus of the stria terminalis (BNST), well connected to the amygdala, and hypothalamic structures such as the paraventricular (PVH) and dorsomedial (DMH), the arcuate (ARH) nuclei and the lateral hypothalamic areas (LHA) known to control feeding and motivated behaviors. We particularly focused our study on afferences to proopiomelanocortin (POMC), agouti-related peptide (AgRP), melanin-concentrating-hormone (MCH) and orexin (ORX) neurons characteristics of the ARH and the LHA, respectively. We found light to intense innervation of all these hypothalamic nuclei. We particularly showed an innervation of POMC, AgRP, MCH and ORX neurons by the dorsomedial and dorsolateral divisions of the BNST. Therefore, these results lay the foundation for a better understanding of the neuroanatomical basis of the stress-related feeding behaviors.

Dietary Supplementation With Various Fat Oils Affect Phytohemagglutinin Skin Test in Broiler Chickens.

The current study aimed to investigate the effect of different dietary supplemental oils on the immune status of broilers. One-day-old Cobb 500 broiler chicks were randomly distributed into eight batteries and fed eight experimental diets. There were 680 broilers, 85 birds per battery. The experimental oils were all used at 10% of the total diet. Each dietary treatment (TRT) contained one of the following essential oils: TRT 1 = control group that received a basal diet + soybean oil (SO); TRT 2 = basal diet as in TRT 1 + sunflower oil (SFO); TRT 3 = basal diet as in TRT 1 + canola oil (CO); TRT 4 = basal diet as in TRT 1 + flaxseed oil (FLO); TRT 5 = basal diet as in TRT 1 + fish oil (FO); TRT 6 = basal diet as in TRT 1 + mix of fish oil and soya oil (SO + FO); TRT 7 = basal diet as in TRT 1 + algal biomass oil (DHA); TRT 8 = basal diet as in TRT 1 + echium oil (EO). All samples were taken from 10 birds per treatment (n = 10). The immune parameters investigated involved measurement of weights of immune organs as a general indicator, hemocytometric measurements, intestinal microbial count and hindgut acidosis, hindgut volatile fatty acids, and cellular immune response using phytohemagglutinin test. The use of the different dietary treatments did not affect the general health status of the chickens, and the mortality was minimal with no signs of illness or outbreaks. The fact that both the control and the treatment diets were equally consumed would indicate that supplemental oil inclusions did not adversely affect the palatability of the diet by the chickens. At 3 weeks of age, there was no significant effect observed in the microbial counts of the intestine. However, at 5 weeks of age, the highest microbial count was significantly observed for broilers fed EO (7.30%), closely followed by SFO (6.95%), and the least microbial counts were observed for CO (5.63%). No significance was observed for lactic acid bacteria (LAB) and Salmonella. There was no significance observed for the effect of the dietary treatments on the hindgut volatile acid in the broilers. Wattle swelling changes were significant between dietary treatments. The results revealed that dietary FLO, FO, and DHA oils induced higher cellular response than the other treatments (P = 0.035), representing higher cellular response in these groups. In conclusion, supplemental oils rich in n-3 fatty acids may enhance the immune response in broiler chickens, represented by the intestinal microbial counts and the cellular immune response.

Evaluation of the anti-oxidant and anti-inflammatory effects of the methanolic extract of Ferula szowitsiana root on PHA-induced inflammation in human lymphocytes.

Inflammation is defined as a defensive response of the body against either the endogenous or exogenous triggers, while this process becomes chronic leading to various disorders such as asthma, cancers, and multiple sclerosis. Recently, pharmacological properties of different constituents of F. szowitsiana have been reported. In the present study, we aimed to evaluate the anti-oxidative and anti-inflammatory effects of the methanolic extract of F. szowitsiana root on human isolated lymphocytes. The effects of either F. szowitsiana (10, 40 and 160 µg/ml) or dexamethasone (0.1 mM) were evaluated on the levels of cell proliferation, reactive oxygen species (ROS), nitric oxide (NO) production, malondialdehyde (MDA), superoxide dismutase (SOD) and catalase (CAT) activities, and total glutathione content (GSH) as well as the secretion of inflammatory cytokines [interleukin 6 (IL-6) and tumor necrosis factor (TNF)-α] in the presence or absence of phytohemagglutinin (PHA) stimulation (n = 8 for each group). PHA stimulation notably elevated ROS, NO, MDA, IL-6, and TNF-α levels as well as diminished GSH, CAT and SOD levels. In PHA-stimulated, the results also revealed that F. szowitsiana (10-160 µg/ml) significantly decreased MDA, ROS, NO, IL-6 and TNF-α levels as well as increased CAT, SOD and GSH levels. Collectively, F. szowitsiana is able to attenuate the overproduction of inflammatory and oxidative stress markers in the presence of PHA-stimulated T lymphocytes, while to propagate the anti-oxidative defense. Contextually, the plant has promising healing effects in the different inflammatory disorders associated with the interference of the acquired immune system such as multiple sclerosis and asthma.

Astrocaryum aculeatum fruit improves inflammation and redox balance in phytohemagglutinin-stimulated macrophages.

ETHNOPHARMACOLOGICAL RELEVANCE: The fruit of Astrocaryum aculeatum G.Mey. (tucumã) is highly consumed by riverside communities in the Amazonian region. These communities have recently been shown to have increased longevity and reduced prevalence of age-related morbidity. Tucumã, which is locally used in their diet and traditional medicine may contribute to these features. AIM OF THE STUDY: To investigate the anti-inflammatory and antioxidant properties of A. aculeatum extract against phytohemagglutinin-induced inflammation in cell cultures. MATERIALS AND METHODS: Cell viability and cytotoxicity assays, gene expression of interleukins IL-1ß, IL-6, IL-10, levels of reactive oxygen species (ROS), nitric oxide (NO) and thiols were employed, as well as the activities of antioxidant enzymes in RAW 264.7 cells stimulated with phytohemagglutinin to mimic inflammation. RESULTS: The extract of A. aculeatum fruit inhibited macrophage proliferation (P < 0.05), arrested the cell cycle in G0/G1 phase (P < 0.001), increased antioxidant defenses (P < 0.01), reduced oxidative stress (P < 0.01), and modulated genes related to the inflammatory response (P < 0.001). CONCLUSION: Our results demonstrate that A. aculeatum fruit has anti-inflammatory and antioxidant capacities. These beneficial effects of tucumã on cells are also likely to be seen in vivo, thereby suggesting that its extract is a suitable therapeutic adjuvant in the prevention or treatment of inflammatory diseases.

Mitogen-Induced Interferon Gamma Production in Human Whole Blood: The Effect of Heat and Cations.

BACKGROUND: Interferon-gamma release assays (IGRAs) are blood tests used to measure the amount of interferon-γ (IFN-γ) released by T lymphocytes after stimulation by antigens specific for the diagnosis of latent tuberculosis infection. A mitogen serves as a positive control to assess the immune function in IGRAs. METHODS: This in vitro study was conducted to evaluate IFN-γ production by human whole blood stimulated with heat-treated and/or cation-supplemented phytohemagglutinin (PHA), concanavalin A (Con A) and pokeweed mitogen (PWM), using QuantiFERON-TB Gold Kit ELISA tests. RESULTS: The optimal concentrations of PWM, Con A and PHA for IGRAs were 2 µg/mL, 5 µg/mL and 10 µg/mL, respectively. The results showed that IFN-γ production in response to PWM was the highest and PHA was the lowest amount. The median values of three mitogens were in the following order: PWM≥Con A≥ positive control>>PHA-P>>negative control. PWM and PHA were heat stable, while Con A was heat sensitive. The mitogen response of lymphocytes to untreated or heat-treated PWM and heat-treated Con A was increased in 1 mM Ca2+-supplemented groups, whereas the response to heat-treated PHA was decreased. Exposure to 1 mM Mg2+ had no effect on untreated or heat-treated PWM, and a concentration of 1 mM Zn2+ inhibited the stimulation of un-treated PWM. We found that calcium supplementation improved the PWM-induced production of IFN-γ. CONCLUSION: Therefore, PWM is an appropriate mitogen for use as a positive control in IGRAs. It is a potential indicator of cytokine production in the diagnostic as well as research settings, and calcium supplementation improved stimulation.

Do glucocorticoids or carotenoids mediate plumage coloration in parrots? An experiment in Platycercus elegans.

Conspicuous coloration can indicate phenotypic quality, and may reflect exposure or vulnerability to stress, or access to essential nutrients such as pigments. Although the production of pigmented colours is well understood, much less is known about how structural colours are affected by physiological state. In this study, we tested whether glucocorticoids (corticosterone) predicted expression of plumage coloration in an Australian parrot, the crimson rosella (Platycercus elegans). Parrots provide an interesting and unique test, as they possess conspicuous coloration produced by distinctive pigments known as psittacofulvins, in addition to structural coloration. We have previously documented that coloration in P. elegans is condition-dependent and responds to dietary manipulation. Here, n = 21 P. elegans underwent a dietary manipulation (including food restriction or carotenoid supplementation) during which they moulted, and the change in reflectance was measured for three structural and three pigmentary plumage patches. Stress-induced corticosterone (10 min after handling) measured at the start of the experiment predicted change in coloration in two pigmentary patches (crown and front). We also found that change in stress-induced corticosterone during the experiment was associated with the change in coloration of the crown and two structural patches (cheek and epaulette). Baseline corticosterone (<3 min after handling) was not associated with any measure of coloration. We found no effects of dietary manipulation on baseline or stress-induced corticosterone, but carotenoid supplementation was associated with an increase in a measure of chronic stress (heterophil/lymphocyte ratio), and the corticosterone response to handling decreased over the course of the study. Our results suggest that corticosterone may be linked to colour expression more broadly than previously recognised, including psittacofulvin and structural coloration in parrots, and they confirm the independence of plumage pigmentation in parrots from carotenoid accumulation. Moreover, our study provides new insight into the stress responses of Psittaciformes, one of the most highly threatened avian orders.

Inhibitory effect of berberine on interleukin-2 secretion from PHA-treated lymphocytic Jurkat cells.

Berberine is an isoquinoline alkaloid isolated from herb plants, such as Cortex phellodendri (Huangbai) and Rhizoma coptidis (Huanglian). Huanglian and Huangbai have been used as "heat-removing" agents. In addition, berberine has been reported to exert anti-inflammatory effect both in vivo and in vitro, where mitogen-activated protein kinase (MAPK) and cyclooxygenase-2 (COX-2) expressions are critically implicated. We herein tested the hypothesis that berberine exerts an anti-inflammatory effect through MAPK and COX-2 signaling pathway in T-cell acute lymphoblastic leukemia (T-ALL). In Jurkat cells, we found that PHA exposure caused elevation on interleukin-2 (IL-2) production in a time-dependent manner. PHA-stimulated reactions were steeply suppressed by berberine, such as IL-2 mRNA expression and protein secretion. However, berberine did not exert any cytotoxic effect at doses of 40 µg/ml. In addition, the possible molecular mechanism of anti-inflammation effect of berberine could be the inhibition of PHA-evoked phosphorylation of p38, since c-Jun N-terminal kinases (JNK) and extracellular signal-regulated kinase (ERK) expressions did not alter. Consistent with above results, berberine inhibition on PHA-induced IL-2 secretion could be reversed by treatment of SB203580, a specific inhibitor of p38-MAPK. Interestingly, upregulation of PHA-induced COX-2 expression was also observed following berberine treatment of Jurkat cells. Furthermore, flow cytometry analysis showed berberine-induced cell cycle arrest at G1 phase after PHA stimulation and decreased percentage of G2/M phase. In conclusion, our study demonstrated that the anti-inflammatory effect of berberine largely potentially results from its ability to attenuate p38 MAPK expression, and does not exclude a positive action of berberine on cell cycle arrest. These results provide an innovative medicine strategy to against or treat T-ALL patients.

Verification of the anti-inflammatory activity of the polyphenolic-rich fraction of Araucaria bidwillii Hook. using phytohaemagglutinin-stimulated human peripheral blood mononuclear cells and virtual screening.

ETHNOPHARMACOLOGICAL RELEVANCE: Araucaria bidwillii Hook., the bunya pine, is an evergreen plant belonging to family Araucariaceae. Traditionally, its leaves and oleoresins have been used as herbal remedies to alleviate pain and inflammation. Based upon the frequent adverse effects accompanying synthetic anti-inflammatory drugs, this study will assess the anti-inflammatory activity of both the total methanol extract and the polyphenolic-rich fraction of A. bidwillii leaves. MATERIALS AND METHODS: The anti-inflammatory activity was assessed in vitro using phytohaemagglutinin-stimulated human peripheral blood mononuclear cells (PBMCs). Isolation of the major compounds was conducted using various chromatographic techniques. Molecular modelling studies are performed on tumor necrosis factor-α (TNF-α), cyclooxygenase-II (COX-II) and 5-lipooxygenase (5-LOX). RESULTS: Both the total methanol extract of Araucaria bidwillii leaves and its fraction revealed a dose-dependent manner in lowering the levels of IL-1ß, IL-6 and TNF-α with an equivalent activity to that of indomethacin. In addition, the phytochemical investigation of the polyphenolic-rich fraction results in identification of agathisflavone-4',7''-dimethyl ether (1), 7-O-methyl-6-hydroxyapigenin (2) and 4',4'-di-O-methylamentoflavone (3) as the main components. In silico molecular modelling showed that agathisflavone-4',7''-dimethyl ether (1) exhibited the fittest binding in TNF-α active sites, while 7-O-methyl-6-hydroxyapigenin (2) showed the highest inhibition in COX-II whereas 4',4'-di-O-methylamentoflavone (3) is the most potent 5-LOX inhibitor. CONCLUSION: Thus, the leaves of Araucaria bidwillii could afford a potent anti-inflammatory agent that effectively ameliorates inflammation and its related hazards. This in turn consolidates the fact of using the leaves of Araucaria bidwillii to sooth inflammation in traditional medicine.

Javamide-II Found in Coffee Is Better than Caffeine at Suppressing TNF-α Production in PMA/PHA-Treated Lymphocytic Jurkat Cells.

Recent studies have suggested positive benefits of coffee consumption on inflammation-related diseases, such as liver diseases and diabetes, where activated lymphocytes and TNF-α are critically implicated. Interestingly, some reports suggested that javamide-II found in coffee may have anti-inflammatory activity greater than that of caffeine, but there is limited information about its effect on TNF-α production by activated lymphocytes. Therefore, the inhibitory effect of javamide-II on TNF-α was investigated in PMA/PHA-treated lymphocytic Jurkat cells. At 5 µM, javamide-II, not caffeine, inhibited TNF-α production in the cells (45 ± 4%, P < 0.001). To elucidate the underlying mechanism, the phosphorylation of MAP kinases (ERK, p38, and JNK) was investigated in the Jurkat cells. Javamide-II had little effect on JNK or p38 phosphorylation, but javamide-II (<20 µM) decreased ERK phosphorylation, consequently reducing TNF-α mRNA expression in the cells ( P < 0.001). The involvement of ERK phosphorylation was also confirmed by an ERK1/2 inhibitor (SCH772984). Furthermore, javamide-II was also found to inhibit IL-2 production, which is up-regulated by ERK phosphorylation in cells ( P < 0.001). These data suggested that javamide-II may be a potent compound to suppress TNF-α production more efficiently than caffeine by inhibiting ERK phosphorylation in Jurkat cells.

Auraptene regulates Th1/Th2/TReg balances, NF-κB nuclear localization and nitric oxide production in normal and Th2 provoked situations in human isolated lymphocytes.

BACKGROUND: Auraptene as member of dietary coumarins, is found in citrus fruits. Former studies have demonstrated its anti-inflammatory and anti-cancer activity. PURPOSE: The mechanism of action and immune-modulatory property of this compound on human lymphocytes are greatly unknown. STUDY DESIGN/METHODS: The effect of three concentrations (10, 30 and 90 µM) of auraptene or dexamethasone (0.1 mM) were evaluated on percentage of cell proliferation and nitric oxide (NO) production as well as secretion and gene expression of cytokines, and NF-κB level in PHA-stimulated and non-stimulated lymphocytes. RESULTS: In non-stimulated cells, all three concentrations of auraptene significantly increased the gene expression index of IL-10 (P < 0.05-0.001). The IFN-γ gene expression index, IFN-γ/IL-4 and IL-10/IL-4 gene expression ratio were significantly increased due to the high concentration (90 µM) of auraptene treatment compared to control group (P < 0.05-0.001). In PHA stimulation, all three concentrations of the extract significantly decreased proliferation, cytokines (IL-4, IL-10 and IFN-γ) and NF-κB level as well as NO production, but IFN-γ/IL-4 and IL-10/IL-4 ratio were significantly increased compared control group (P < 0.05-0.001). Gene expression of IL-10 and IL-4 was decreased but that of IFN-γ as well as FN-γ/IL-4 and IL-10/IL-4 ratio were significantly increased due to all three concentrations of auraptene. CONCLUSION: The results showed promoting effects of auraptene on T cell subsets toward Th1 (IFN-γ) and Treg (IL-10), which suggest its therapeutic value for treatment of Th2 cells predominant diseases including allergic disease such as asthma and atopic dermatitis as well as cancers.

Efferents of anterior cingulate areas 24a and 24b and midcingulate areas 24a' and 24b' in the mouse.

The anterior cingulate cortex (ACC), constituted by areas 25, 32, 24a and 24b in rodents, plays a major role in cognition, emotion and pain. In a previous study, we described the afferents of areas 24a and 24b and those of areas 24a' and 24b' of midcingulate cortex (MCC) in mice and highlighted some density differences among cingulate inputs (Fillinger et al., Brain Struct Funct 222:1509-1532, 2017). To complete this connectome, we analyzed here the efferents of ACC and MCC by injecting anterograde tracers in areas 24a/24b of ACC and 24a'/24b' of MCC. Our results reveal a common projections pattern from both ACC and MCC, targeting the cortical mantle (intracingulate, retrosplenial and parietal associative cortex), the non-cortical basal forebrain, (dorsal striatum, septum, claustrum, basolateral amygdala), the hypothalamus (anterior, lateral, posterior), the thalamus (anterior, laterodorsal, ventral, mediodorsal, midline and intralaminar nuclei), the brainstem (periaqueductal gray, superior colliculus, pontomesencephalic reticular formation, pontine nuclei, tegmental nuclei) and the spinal cord. In addition to an overall denser ACC projection pattern compared to MCC, our analysis revealed clear differences in the density and topography of efferents between ACC and MCC, as well as between dorsal (24b/24b') and ventral (24a/24a') areas, suggesting a common functionality of these two cingulate regions supplemented by specific roles of each area. These results provide a detailed analysis of the efferents of the mouse areas 24a/24b and 24a'/24b' and achieve the description of the cingulate connectome, which bring the anatomical basis necessary to address the roles of ACC and MCC in mice.

Long-term sex-differential effects of neonatal vitamin A supplementation on in vitro cytokine responses.

High-dose vitamin A supplementation (VAS) may affect mortality to infectious diseases in a sex-differential manner. Here, we analysed the long-term immunological effects of neonatal vitamin A supplementation (NVAS) in 247 children, who had been randomly allocated to 50 000 or 25 000 IU vitamin A (15mg and 7·5mg retinol equivalents, respectively) or placebo at birth. At 4-6 months of age, we assessed bacille Calmette-Guérin (BCG) scarification, and we analysed in vitro responses of TNF-α, IL-5, IL-10, IL-13 and IFN-γ in whole blood stimulations to phytohaemagglutinin (PHA), purified protein derivative (PPD), tetanus toxoid and lipopolysaccharide. There were no differences between the two doses of NVAS, and thus they were analysed combined as NVAS (any dose) v. placebo. All analyses were performed unstratified and by sex. NVAS increased the chance of having a scar after BCG vaccination in females (NVAS v. placebo: 96 v. 71 %, proportion ratio: 1·24; 95 % CI 1·09, 1·42), but not in males (P for interaction=0·012). NVAS was associated with significant sex-differential effects on the pro- to anti-inflammatory cytokine ratios (TNF-α:IL-10) to PPD, tetanus toxoid and medium alone, which were increased in females but decreased in males. In addition, IL-17 responses tended to be increased in NVAS v. placebo recipients in males but not in females, significantly so for the PHA stimulation. The study corroborates sex-differential effects of VAS on the immune system, emphasising the importance of analysing VAS effects by sex.

Immune responses, intestinal microbiota, performance and blood characteristics of Japanese quail fed on diets containing camphor.

This study aimed to investigate the effect of supplemental camphor on the performance and immune functions of Japanese quail by feeding graded levels (0 (control), 250, 500, 750, 1000, 5000 or 10 000 ppm) of camphor during a 42-day feeding trial. In all, 280 1-day-old quail chicks were randomly assigned into 28 cages of 10 chicks each with separate feeders. The results clearly demonstrated that camphor did not have a significant effect on BW, BW gain, total experimental average daily feed intake, feed conversion ratio, internal organ relative weights and biochemical parameters such as uric acid, albumin, total protein and triglyceride; however, plasma cholesterol concentration was significantly different in a linear manner, in which 500 ppm of camphor resulted in a lower level of cholesterol. Alternatively, greater concentrations of glucose and high-density lipoprotein (HDL) were also found in 5000 and 1000 ppm of camphor groups, respectively. Cellular responses to the phytohaemagglutinin-P and 2,4-dinitro 1-chlorobenzene skin test were not influenced by dietary camphor. Humoral responses to secondary sheep red blood cells, avian influenza virus (AIV) and Newcastle disease virus (NDV) immunisations were positively influenced by camphor supplementation, in which greater secondary response to sheep erythrocytes belonged to 750 and 1000 ppm of camphor groups; whereas, diet supplementation with camphor had no significant effect on lymphoid organ weights and heterophil-to-lymphocyte ratio. The greatest AIV antibody titers were seen in groups, which received 1000 and 5000 ppm of camphor (P<0.05) and the values of NDV antibody titers increased with an increase in the camphor consumption. Furthermore, dietary inclusion of 500 ppm of camphor positively decreased coliform populations in the gastrointestinal tract (GIT). In addition, an increase in lactic acid bacteria was also observed in quails, which were fed on the diets containing 750 ppm camphor. Collectively, these data suggest that as a phytogenic feed additive, camphor may effectively act as a modulator of health status (by increasing glucose and HDL), GIT microbiota and immunological responses of the Japanese quail.

Tableta masticable de lecitina de soya: de subproducto a producto farmacéutico / Chewable pill of soy phosphatidylcholine: from by-product to pharmaceutical product

En el Laboratorio Farmacéutico Oriente de Santiago de Cuba se acometió el desarrollo de una tableta masticable de lecitina de soya con fines de registro y ulterior producción, lo cual se realizó durante el bienio 2011-2013. Se utilizaron excipientes de calidad farmacéutica, los métodos analíticos de la Farmacopea de los Estados Unidos, edición 35/Formulario Nacional, edición 30 del 2012, así como la tecnología de granulación húmeda y compresión directa. La lecitina fue caracterizada como materia prima farmacéutica y la tableta desarrollada cumplió con los atributos de calidad establecidos, por lo cual se registró con estabilidad comprobada de 2 años. Se suministró valor agregado a esta sustancia, con riesgo potencial de acumulación para el medio ambiente, como producto farmacéutico nuevo en Cuba(AU)

The development of a chewable pill of soy phosphatidylcholilne was undertaken in Oriente Pharmaceutical Laboratory from Santiago de Cuba with registration ends and subsequent production, that was carried out during the biennium 2011-2013. Excipients of pharmaceutical quality, the analytic methods of the United States Pharmacopoeia, edition 35/National Form, 2012 30th edition, as well as the technology of humid granulation and direct compression were used. Phosphatidylcholine was characterized as pharmaceutical raw material and the developed pill fulfilled the established quality attributes, reason why it was registered with 2 years proven stability. Added value was given to this substance, with potential risk of accumulation for the environment, as new pharmaceutical product in Cuba(AU)

Tableta masticable de lecitina de soya: de subproducto a producto farmacéutico / Chewable pill of soy phosphatidylcholine: from by-product to pharmaceutical product

En el Laboratorio Farmacéutico Oriente de Santiago de Cuba se acometió el desarrollo de una tableta masticable de lecitina de soya con fines de registro y ulterior producción, lo cual se realizó durante el bienio 2011-2013. Se utilizaron excipientes de calidad farmacéutica, los métodos analíticos de la Farmacopea de los Estados Unidos, edición 35/Formulario Nacional, edición 30 del 2012, así como la tecnología de granulación húmeda y compresión directa. La lecitina fue caracterizada como materia prima farmacéutica y la tableta desarrollada cumplió con los atributos de calidad establecidos, por lo cual se registró con estabilidad comprobada de 2 años. Se suministró valor agregado a esta sustancia, con riesgo potencial de acumulación para el medio ambiente, como producto farmacéutico nuevo en Cuba

The development of a chewable pill of soy phosphatidylcholilne was undertaken in Oriente Pharmaceutical Laboratory from Santiago de Cuba with registration ends and subsequent production, that was carried out during the biennium 2011-2013. Excipients of pharmaceutical quality, the analytic methods of the United States Pharmacopoeia, edition 35/National Form, 2012 30th edition, as well as the technology of humid granulation and direct compression were used. Phosphatidylcholine was characterized as pharmaceutical raw material and the developed pill fulfilled the established quality attributes, reason why it was registered with 2 years proven stability. Added value was given to this substance, with potential risk of accumulation for the environment, as new pharmaceutical product in Cuba

Aspectos farmacológicos de la lecitina de soya y sus posibles aplicaciones médicas / Pharmacological aspects of soy phosphatidylcholine and its possible medical uses

La lecitina de soya, producto natural empleado como suplemento nutricional, presenta múltiples acciones biológicas demostradas, por lo cual resulta muy beneficiosa para tratar a pacientes con distintas afecciones. Teniendo en cuenta lo anterior se realizó la presente investigación donde se exponen algunos aspectos de interés, con vistas a difundir aún más lo relacionado con esta temática(AU)

The soy phosphatidylcholine, natural product used as nutritional supplement, presents multiple demonstrated biological actions, reason why it is very beneficial to treat patients with different disorders. Taking into account the above-mentioned the present investigation was carried out where some aspects of interest are exposed, aimed at diffusing even more everything related to this thematic(AU)