Phytochemicals mediated modulation of microRNAs and long non-coding RNAs in cancer prevention and therapy.

MicroRNAs (miRNAs) and long noncoding RNAs (lncRNAs) are two main categories of noncoding RNAs (ncRNAs) that can influence essential biological functions in various ways, as well as their expression and function are tightly regulated in physiological homeostasis. Additionally, the dysregulation of these ncRNAs seems to be crucial to the pathogenesis of human diseases. The latest findings indicate that ncRNAs execute vital roles in cancer initiation and progression, and the cancer phenotype can be reversed by modulating their expression. Available scientific discoveries suggest that phytochemicals such as polyphenols, alkaloids, terpenoids, and organosulfur compounds can significantly modulate multiple cancer-associated miRNAs and lncRNAs, thereby inhibiting cancer initiation and development. However, despite promising outcomes of experimental research, only a few clinical trials are currently being conducted to evaluate the therapeutic effectiveness of these compounds. Nevertheless, understanding phytochemical-mediated ncRNA regulation in cancer and the underlying molecular mechanisms on tumor pathophysiology can aid in the development of novel therapeutic strategies to combat this deadly disease.

Roles of microRNAs in chronic pediatric diseases and their use as potential biomarkers: A review.

MicroRNAs (miRNAs) are small non-coding highly conserved RNA molecules that can act as master regulators of gene expression in a sequence-specific manner either by translation repression or mRNA degradation, influencing a wide range of biologic processes that are essential for the maintenance of cellular homeostasis. Chronic pediatric diseases are the leading cause of death worldwide among children and the recent evidence indicates that aberrant miRNA expression significantly contributes to the development of chronic pediatric diseases. This review focuses on the role of miRNAs in five major chronic pediatric diseases including bronchial asthma, congenital heart diseases, cystic fibrosis, type 1 diabetes mellitus, and epilepsy, and their potential use as novel biomarkers for the diagnosis and prognosis of these disorders.

Creation of optical speckle by randomizing a vortex-lattice.

We investigate the number of vortices embedded in a carrier beam needed to produce a speckle pattern and the necessary conditions in terms of their initial distribution and topological charges. A spatial light modulator is used to imprint arrays of vortices in a Gaussian beam, which is propagated in free space for a given distance and then focused in order to induce interaction among the vortices in the focal region. The resulting optical field is analyzed after propagation up to a transverse plane where the carrier beam would recover its initial size in the absence of vortices. The role of different control parameters for obtaining ordered and disordered patterns is discussed. Our experimental study is complemented with a thorough numerical analysis, from which the statistical properties of the disordered patterns are characterized, and the conditions for obtaining well-developed speckle are determined. We also discuss the creation and annihilation of vortex pairs, depending on the initial conditions.

Energy and nitrogen partitioning in dairy cows at low or high metabolizable protein levels is affected differently by postrumen glucogenic and lipogenic substrates.

This study tested the effects of energy from glucogenic (glucose; GG) or lipogenic (palm olein; LG) substrates at low (LMP) and high (HMP) metabolizable protein levels on whole-body energy and N partitioning of dairy cattle. Six rumen-fistulated, second-lactation Holstein-Friesian dairy cows (97 ± 13 d in milk) were randomly assigned to a 6 × 6 Latin square design in which each experimental period consisted of 5 d of continuous abomasal infusion followed by 2 d of rest. A total mixed ration consisting of 42% corn silage, 31% grass silage, and 27% concentrate (dry matter basis) was formulated to meet 100 and 83% of net energy and metabolizable protein requirements, respectively, and was fed at 90% of ad libitum intake by individual cow. Abomasal infusion treatments were saline (LMP-C), isoenergetic infusions (digestible energy basis) of 1,319 g/d of glucose (LMP-GG), 676 g/d of palm olein (LMP-LG; major fatty acid constituents are palmitic, oleic, and linoleic acid), or 844 g/d of essential AA (HMP-C), or isoenergetic infusions of 1,319 g/d of glucose + 844 g/d of essential AA (HMP-GG) or 676 g/d of palm olein + 844 g/d of essential AA (HMP-LG). The experiment was conducted in climate respiration chambers to determine energy and N balance in conjunction with milk production and composition, nutrient digestibility, and plasma constituents. Infusion of GG and LG decreased dry matter intake, but total gross energy intake from the diet plus infusions was not affected by GG or LG. Furthermore, GG or LG did not affect total milk, protein, or lactose yields. Infusing GG or LG at the HMP level did not affect milk production differently than at the LMP level. Infusion of GG stimulated energy retention in body tissue, increased plasma glucose and insulin concentrations, decreased lipogenic metabolites in plasma, and decreased milk fat yield and milk energy output. Nitrogen intake decreased and milk N efficiency increased in response to GG, and N retention was not affected. Infusion of LG tended to increase metabolizable energy intake, increased milk fat yield and milk energy output, increased plasma triacylglycerides and long-chain fatty acid concentrations, and had no effect on energy retention. Infusion of LG decreased N intake but did not affect milk N efficiency or N retention. Compared with the LMP level, the HMP level increased dry matter intake, gross and metabolizable energy intake, and total milk, fat, protein, and lactose yields. Milk energy output increased at the HMP level, and protein level did not affect total energy retention. Heat production increased at the HMP level, but only when GG and LG were infused. The HMP level increased N intake, milk N output, and plasma urea concentration, tended to increase N retention, and decreased milk N efficiency. Regardless of protein level, GG promoted energy retention and improved milk N efficiency, but not through increased milk protein yield. Infusion of LG partitioned extra energy intake into milk and had no effect on milk N efficiency.

Feed and nitrogen efficiency are affected differently but milk lactose production is stimulated equally when isoenergetic protein and fat is supplemented in lactating dairy cow diets.

Fifty-six Holstein-Friesian cows were used in a randomized complete block design to test the effects of supplemental energy from protein (PT) and fat (FT) on lactation performance and nutrient digestibility in a 2 × 2 factorial arrangement. During the control period, cows were adapted for 28 d to a basal total mixed ration consisting of 34% grass silage, 33% corn silage, 5% grass hay, and 28% concentrate on a dry matter (DM) basis. Experimental rations were fed for 28 d immediately following the control period and consisted of (1) low protein, low fat (LP/LF), (2) high protein, low fat (HP/LF), (3) low protein, high fat (LP/HF), or (4) high protein and high fat (HP/HF). To obtain the HP and HF diets, intake of the basal ration was restricted and supplemented isoenergetically (net energy basis) with 2.0 kg/d of rumen-protected protein (soybean + rapeseed, 50:50 mixture on DM basis) and 0.68 kg/d of hydrogenated palm fatty acids (FA) on a DM basis. Milk production and composition, nutrient intake, and apparent digestibility were measured during the final 7 d of the control and experimental periods. No interaction was found between PT and FT on milk production and composition. Yields of milk, fat- and protein-corrected milk, and lactose increased in response to PT and FT and lactose concentration was unaffected by treatment. Milk protein concentration and yield increased in response to PT, and protein yield tended to increase in response to FT. Milk fat concentration and yield increased in response to FT and were unaffected by PT. Milk urea concentration increased and nitrogen efficiency decreased in response to PT. Feed and nitrogen efficiency were highest on the LP/HF diet and both parameters increased in response to FT, whereas milk urea concentration was not affected by FT. Energy from fat increased the concentration and yield of ≥16-carbon FA in milk and decreased the concentration of FA synthesized de novo, but had no effect on their yield. Concentration and yield of de novo-synthesized FA increased in response to PT. Concentration and yield of polyunsaturated FA increased and decreased in response to PT and FT, respectively. Apparent total-tract digestibility of crude fat decreased in response to PT, and FT increased crude protein digestibility. Energy supplementation through rumen-inert hydrogenated palm FA appears to be an efficient feeding strategy to stimulate milk production with regard to feed and nitrogen efficiency compared with supplementing an isoenergetic level of rumen-protected protein.

Regulatory mechanisms of microRNAs in endocrine disorders and their therapeutic potential.

MicroRNAs (miRNAs) are small endogenous non-coding RNA molecules capable of regulating gene expression at the post-transcriptional level either by translational inhibition or mRNA degradation and have recently been importantly related to the diagnosis and prognosis of the most relevant endocrine disorders. The endocrine system comprises various highly vascularized ductless organs regulating metabolism, growth and development, and sexual function. Endocrine disorders constitute the fifth principal cause of death worldwide, and they are considered a significant public health problem due to their long-term effects and negative impact on the patient's quality of life. Over the last few years, miRNAs have been discovered to regulate various biological processes associated with endocrine disorders, which could be advantageous in developing new diagnostic and therapeutic tools. The present review aims to provide an overview of the most recent and significant information regarding the regulatory mechanism of miRNAs during the development of the most relevant endocrine disorders, including diabetes mellitus, thyroid diseases, osteoporosis, pituitary tumors, Cushing's syndrome, adrenal insufficiency and multiple endocrine neoplasia, and their potential implications as disease biomarkers.

High mortality due to gastrointestinal TB in HIV and non-HIV patients.

BACKGROUND: Due to the reported low incidence of gastrointestinal TB, there is a lack of data related to the prognosis, risk factors and frequency of resistant TB in this subgroup of patients.OBJECTIVE: To report the clinical presentation, diagnostic methods, treatment and outcomes in gastrointestinal TB.METHODS: We prospectively studied the demographic, clinical, and paraclinical data of all consecutive gastrointestinal TB inpatients over an 8-year period.RESULTS: We identified gastrointestinal TB in 28 (3.5%) out of 799 inpatients with TB infection. Seven patients (25%) were HIV-positive. Overall mortality was 35.7%, with the combined variable of haemoglobin <12 g/dL and albumin <2.8 g/dL being independently associated with mortality (OR 25.7, 95% CI 1.405-471.1, P = 0.029). No difference in the need for surgery (28.6% vs. 47.6%, P = 0.662), occurrence of septic shock (14.3 vs. 23.8%, P = 1.00) or mortality (14.3% vs. 42.9%, P = 0.364) was found between HIV and non-HIV patients.CONCLUSION: Gastrointestinal TB was rare among TB patients in Hospital Universitario "Dr José E. González" (3.5%), but had a high mortality rate (35.7%). Clinical evolution, drug susceptibility patterns and outcomes were similar in HIV and non-HIV patients. In both groups, the combined haemoglobin and albumin variable on admission was clearly associated with mortality.

Determination of Sarcocystis lamacanis microcysts in the cardiac muscle of alpacas (Vicugna pacos) and their correlation with troponin cTnI. A study performed in the high Andean region of southern Peru.

The breeding of alpaca (Vicugna pacos) is one of the most important economic activities in the high Andean areas of Peru. The commercialization of products derived from alpaca represents more than 80% of the income of high-Andean families. However, the infestation of parasites such as Sarcocystis lamacanis in the alpacas causes economic losses that deteriorate the already diminished quality of life of the alpaca breeder. The search for biomarkers that allow the early detection of these parasites is one of the most critical challenges in Peru, a country with the largest population of alpacas worldwide. This work aimed to analyze and quantify the microcysts formed by the parasite and relate them to the troponin cTnI level in the blood serum. Troponins are proteins secreted when there is damage to the cardiac muscle. 60 blood and cardiac tissue samples were collected from Tisco and La Raya slaughterhouses, localities of Caylloma Province in Arequipa, and Chucuito District in Puno, both regions in southern Peru. The cardiac muscle samples were processed with the routine histology technique and stained with hematoxylin and eosin. In addition, serum samples were processed with the ELISA and immunochromatography methods for troponin cTnI. Results were 100% positive for the presence of Sarcocystis lamacanis microcysts in all cardiac muscle samples. The average microcyst quantification per field of 100x were 3.5 and 5.7 for the Tisco and La Raya samples. In addition, several microscopic lesions were observed in the cardiac muscles: microcyst infiltration between muscle fibers, basophilic microcysts with a thick outer membrane and bradyzoites inside, and tissue displacement. On the other hand, all serum blood samples were negative for troponin cTnI, with both methods, ELISA and immunochromatography. For results, we infer troponin cTnI do not can be used as a biomarker for heart damage caused by Sarcocystis lamacanis parasite in alpacas.

Genetic identification of a rabies virus from an insectivorous bat in an urban area of Rio Grande do Sul, Brazil.

In Latin America, hematophagous bats are the main reservoirs of rabies virus (RABV) to livestock, to other mammals and, occasionally, to human. Nonetheless, reports of exposure of human and pets to RABV upon aggression by non-hematophagous bats are increasing, possibly facilitated by the synanthropic habits of these bats. We, herein, report the detection and genetic identification of a RABV recovered from an insectivorous bat found sick in a student housing building at the Federal University of Santa Maria, Southern Brazil. Taxonomic characterization identified the captured bat as a member of the genus Nyctinomops, family Molossidae, the group of insectivorous bats. Brain fragments of the bat were positive for RABV antigens by fluorescent antibody test (FAT) and for sequences of the nucleoprotein (N) gene by RT-PCR. The N amplicon was submitted to nucleotide sequencing and analysis, showing that the consensus sequences (SV 33/19) had high identity with RABV sequences of insectivorous bats deposited in GenBank. At phylogenetic tree, the N gene sequences of SV 33/19 clustered with RABV recovered from Nyctinomops laticaudatus, Molossus molossus, and Tadarida lauticaudata bats, and a part of RABV variant 3, 4, and 6, that correspond to Desmodus rotundus, Tadarida brasiliensis, and Lasiurus cinereus, respectively. Although no direct human or domestic animal exposure has been reported, this case strengthens the need for a continuous rabies vaccination in pets in the surrounding areas, since non-hematophagous bats may serve as source of infection for these animals. These findings also call attention for continuous monitoring of populations of synanthropic bats to avoid/prevent human exposure.

Identification of iron-acquisition proteins of Avibacterium paragallinarum.

When Avibacterium paragallinarum reference strain 0083 (serovar A) was grown in an iron-restricted culture medium, the expression of the 60, 68 and 93 kDa outer membrane proteins increased as compared with normal media. Sera of chickens experimentally infected with Av. paragallinarum recognized these iron-restriction induced proteins, suggesting their expression in vivo. The three outer membrane proteins were identified as transferrin receptor and iron transport proteins by mass spectroscopy and a search in sequence databases. As these proteins have been reported to be regulated by the Fur protein in many bacteria, we investigated, through molecular methods, the presence of the fur gene in Av. paragallinarum. A candidate fur gene of Av. paragallinarum was amplified by polymerase chain reaction using complementary primers to conserved regions of fur gene sequences from members of the Pasteurellaceae family. The nucleotide sequence of the cloned gene, from ATG to TAA stop codon, was 453 base pairs in length and the deduced amino acid sequence showed 94% identity with Fur sequences of Actinobacillus pleuropneumoniae and Haemophilus ducreyi. The Av. paragallinarum deduced Fur protein (17.8 kDa) amino acid sequence contains the N-terminal helix-turn-helix DNA-binding domain and the two iron-binding sites in the C-terminal end, typical of other described Fur proteins. The study of iron-restriction-induced proteins and the mechanism regulating their expression could lead to an understanding of the responses of Av. paragallinarum to survive in an iron-restricted environment on host mucosal surfaces.

Bioactive triterpenoids from the caffeine-rich plants guayusa and maté.

Unlike all other caffeinated plants, guayusa (Ilex guayusa Loes.) and maté (Ilex paraguariensis A. St. Hill) contain high amounts of pentacyclic triterpenoid acids and alcohols. A phytochemical investigation on these plants revealed a similar triterpenoid profile and a content of ursolic acid (0.7-1%) and amyrin esters (up to 0.5%), quite unusual for dietary plants. The major constituent of the amyrin complex from both plants is α-amyrin palmitate (2a), accompanied by lower amounts of its corresponding palmitoleate (2b) and by the corresponding constitutional isomers from the ß-series (3a and 3b, respectively). Ursolic acid (1) was identified as the responsible for the activity of maté and guayusa extracts in the activation of TGR5, a nuclear receptor of relevance for the prevention and management of diabetes and metabolic syndrome because of its involvement in the regulation of energy expenditure and insulin sensitivity.

Antimicrobial activity of coconut oil-in-water emulsion on Staphylococcus epidermidis and Escherichia coli EPEC associated to Candida kefyr.

Candida kefyr has been considered both a food-spoiling agent and a type of yeast with fermentation properties. In this study, the authors have evaluated the antimicrobial activity of a coconut oil-in-water emulsion associated to the presence of C. kefyr. Fresh coconut kernels were used to obtain the coconut oil-in-water emulsion, the sterile coconut oil-in-water emulsion by decantation, and the coconut oil by means of a heating process. Commercial virgin coconut oil was also used. Agar diffusion, minimal inhibitory concentration and minimal bactericidal concentration (MIC/MBC) techniques were employed to evaluate antimicrobial activity against E. coli and S. epidermidis. The C. kefyr isolate was identified and confirmed. Coconut milk-derived fatty acids were characterized by acid index and thin layer chromatography. Scanning electronic microscopy was performed to evaluate the morphology of the microorganisms. Lipase activity of C. kefyr isolate was also detected. Coconut oil-in-water emulsion associated to C. kefyr was active against both bacteria. Thin layer chromatography confirmed the presence of triglycerides and free fatty acids. The acid index showed higher acidity potential for the coconut oil-in-water emulsion. The microscopic images showed antibacterial action through the formation of membrane holes' and demonstrated yeast shape. All the above show new potentials for C. kefyr and coconut oil-in-water emulsion in food technology.

Clostridium difficile infection (CDI) severity and outcome among patients infected with the NAP1/BI/027 strain in a non-epidemic setting.

OBJECTIVE: Determine whether the NAP1 strain identified by polymerase chain reaction (PCR)-based stool assay is correlated with CDI severity and clinical outcomes. METHODS: Medical records of adult patients with positive stool Xpert® Clostridium difficile PCR assay for an initial episode of CDI between January 2012 and January 2013 at a tertiary care hospital in Chicago were reviewed. Two patients diagnosed with CDI caused by a non-NAP1 strain (positive Xpert® C. difficile assay but negative Xpert® C. difficile Epi assay) were included for each patient diagnosed with CDI caused by a NAP1 strain (positive Epi assay). Patient charts were reviewed for markers of severity, risk factors, treatment regimens, and outcomes. RESULTS: Of 494 stool specimens, 90 (18%) that were positive for C. difficile by PCR were positive for NAP1 strain. In total, 37 patients with CDI due to NAP1 were matched with 74 patients with CDI due to non-NAP1 strains. Multivariable model revealed individuals ≥65 years old were 3 times more likely to have NAP1 strain than individuals <65 (P=.02). Residents of a nursing home prior to hospitalization were 10 times more likely to have NAP1 strain than patients residing in their homes (P=.001). More NAP1 cases had a change in treatment from metronidazole to oral vancomycin plus intravenous metronidazole (P=.01). The severity of CDI, incidence of mortality and recurrent CDI were similar between groups. CONCLUSIONS: In a nonepidemic setting, NAP1 strains were more common in older patients and individuals admitted from nursing homes. Identification of NAP1 by PCR of stool specimens was associated in a change of therapy but did not predict worse outcomes. Reporting strain results may not be clinically useful in routine settings.

Resección laparoscópica de metástasis intestinales de melanoma maligno / Laparoscopic resection of malignant melanoma intestinal metastasis

Resumen Objetivos: El melanoma cutáneo presenta un alto potencial metastásico y constituye la fuente extraabdominal más frecuente de lesión del intestino delgado. El diagnóstico de metástasis gastrointestinales es a menudo una expresión de enfermedad avanzada, con una supervivencia media de entre 6 y 9 meses. Materiales y Método: Presentamos el caso de un paciente varón de 63 años diagnosticado de melanoma cutáneo que acudió a urgencias por dolor abdominal y estreñimiento. Se realizó una TC abdominal donde se informó de la existencia de 2 lesiones metastásicas a nivel de intestino delgado que condicionaban oclusión intestinal. Resultados: El paciente fue intervenido quirúrgicamente bajo abordaje laparoscópico con resección de los dos segmentos intestinales afectos y anastomosis intracorpórea. El informe histopatológico confirmó que se trataban de metástasis de melanoma. Discusión: La oclusión intestinal por metástasis de melanoma maligno es muy infrecuente. La cirugía es el tratamiento de elección en pacientes con metástasis intestinales de melanoma. El tratamiento quirúrgico puede mejorar el pronóstico y estaría indicado casos de metástasis únicas o pacientes sintomáticos con intención paliativa. El abordaje mínimamente invasivo ofrece resultados oncológicos similares a la laparotomía.

Aim: Cutaneous melanoma has a high metastatic potential, being the most frequent extra-abdominal source of small bowel metastasis. The diagnosis of gastrointestinal metastases is often an expression of advanced disease, with an average survival of 6-9 months. Materials and Method: We herein present the case of a 63-year-old male patient diagnosed with cutaneous melanoma who arrived to the emergency department complaining of abdominal pain and constipation. An abdominal CT scan was performed, it revealed two metastatic lesions in the small bowel which marked the mechanical obstruction. Results: Patient underwent a laparoscopy and both involved segments were removed. Pathology exam confirmed the diagnosis of melanoma metastases. Discussion: Surgery excision is the treatment of choice in patients with small bowel metastases from melanoma. Surgical management can improve the prognosis and it would be indicated in cases of single metastases or symptomatic patients with a palliative intention. Minimally invasive approach provides similar oncological results as conventional laparotomy. Small bowel obstruction due to metastases of malignant melanoma is extremely rare.

Ulcerative colitis: effect of sulphasalazine, its metabolites and indomethacin on the ability of human colonic mucosa to metabolize prostaglandins in vitro.

1 Homogenates of mucosa from human colon metabolize [3H]-prostaglandin E1 in the presence of nicotinamide adenine dinucleotide to 15-oxo prostaglandin E1 or 15-oxo, 13,14 dihydro prostaglandin E1. 2 Metabolic capacity of tissue from patients with active ulcerative colitis under treatment with sulphasalazine (0.021 +/- 0.004 nmol/Mg protein +/- s.e. mean) did not differ from mucosa of normal patients (0.02 +/- 0.004 nmol/mg protein) during 1 h incubation. 3 Sulphasalazine inhibits prostaglandin E1 metabolism by mucosal homogenates in a dose-dependent manner with an ID50 of 125 microM. Its therapeutically active metabolite, 5-aminosalicylic acid (2.6 mM) was without significant inhibitory activity. 4 Indomethacin inhibits prostaglandin E1 metabolism by colonic mucosa with an ID50 of 388 microM. 5 At present we cannot clearly relate the therapeutic benefit of sulphasalazine and its therapeutically active metabolite, 5-aminosalicylic acid, in ulcerative colitis to their effects on prostaglandin E synthesis or metabolism in vitro.

Intranasal immunization with HIV reverse transcriptase: effect of dose in the induction of helper T cell type 1 and 2 immunity.

HIV reverse transcriptase (RT) is immunologically recognized by the host after natural infection with HIV. To study the immune response to this nonenvelope protein and the effect of the antigen dose and the immunization route on the differential induction of cellular or humoral responses, CB6F1 mice were immunized intranasally with various doses of recombinant RT (0.025-25 microg/dose) in the presence of the mucosal adjuvant cholera toxin (CT). An antigen-specific, dose-dependent, in vitro proliferative response was observed in splenic cells from all mouse groups immunized with RT and CT. Proliferative responses in salivary gland-associated lymph node (SGALN) cells from the same mice were detected only with higher antigen dose immunizations (2.5, 25 microg/dose). IFN-gamma, a Th1-type cytokine, was detected in RT-stimulated culture supernatants from splenic and SGALN cells from all groups of mice immunized with RT and CT in a dose-dependent fashion. IL-4, a Th2-type cytokine, was detected in RT-stimulated culture supernatants from splenic and SGALN cells from mice immunized with higher doses of RT and CT. RT-specific IgG2a, a Th1-type-related antibody, was detected consistently in sera from all animals immunized with RT and CT and was predominant in mice immunized with lower antigen doses (0.025, 0.25 microg/dose). RT-specific IgG1, a Th2-type-related antibody, was detected consistently in mice immunized with higher antigen doses and was predominant in these groups. These studies demonstrate the immunogenicity of recombinant RT and the effect of the antigen dose in the induction of Th1-type and Th2-type immune responses after mucosal immunization.

The lom gene of bacteriophage lambda is involved in Escherichia coli K12 adhesion to human buccal epithelial cells.

The role of the lom gene of bacteriophage lambda in adhesion of Escherichia coli to human buccal epithelial cells (HBC) was studied testing the adherence of lamda lom+ and lambda lom::TnphoA E. coli lysogens. lambda lom+ prophage increased 50% E. coli adhesion. This effect was not observed with lambda lom::TnphoA. These results suggest that the normal Lom protein participates directly in adhesion or regulates the synthesis of other protein(s), which may be involved in adhesion.

Laboratory aspects of immunology.

The field of clinical immunology is advancing rapidly and will continue to grow and benefit from the knowledge gained in different areas of basic immunology. The diagnosis and evaluation of a child with a suspected primary immune defect can be a challenging task that should be done carefully using the best available laboratory resources. Simple laboratory screening tests, available to most pediatricians, can be done initially and will provide an idea of the level of general immune competence. Referral for a more detailed evaluation should be made in selected cases. A basic understanding of the tests used in immune evaluation, as well as the fact that the results may be affected by multiple variables, is important. Of equal significance is the interaction of the physician with the laboratory in which the samples will be analyzed. Proper sample collection and handling as well as prompt processing by the laboratory are crucial. Equally important is the physician's interpretation of results in the context of the pediatric population. Tests like lymphocyte studies, WBC studies, immunoglobulin subclasses, response to antigenic challenge, and certain complement assays require a higher degree of expertise and should only be sent to laboratories with experience in performing them. Physicians need to gain confidence in the power of diagnosis that the immunology laboratory brings to their patients.

Effect of bakuchiol on leukocyte functions and some inflammatory responses in mice.

The effects of bakuchiol, a meroterpenoid isolated from the leaves of Psoralea glandulosa L., on phospholipase A2 (PLA2) activity from different sources, human neutrophil responses, zymosan air pouch and topical inflammation in mice, were investigated. This natural product was a weak inhibitor of secretory and intracellular PLA2 but dose-dependently reduced the formation of LTB4 and TXB2 by human neutrophils and platelet microsomes, respectively. In addition, bakuchiol inhibited degranulation in human neutrophils, whereas superoxide generation was not affected. In mice, bakuchiol decreased cell migration, myeloperoxidase activity and eicosanoid levels in the air pouch inflammation induced by zymosan. After topical administration, this compound was effective as an inhibitor of oedema and myeloperoxidase activity in the 12-O-tetradecanoylphorbol 13-acetate (TPA)-induced ear oedema and significantly reduced the PGE2 content and ear oedema in the arachidonic acid-induced response. Bakuchiol is a natural anti-inflammatory agent able to control leukocytic functions such as eicosanoid production, migration and degranulation in the inflammatory site.