Effect of ß-glucans on rainbow trout (Oncorhynchus mykiss) IgM+ B cells.

ß-glucans are carbohydrates present in the cell wall of many fungi, which are often used as immunostimulants in feeds for farmed species. Their capacity to activate innate immune responses directly acting on innate cell populations has been widely documented in fish. However, whether they can affect the functionality of adaptive immune cells has been scarcely explored. In this context, in the current work, we have determined the effects of ß-glucans on rainbow trout blood IgM+ B cells in the presence or absence of 2,4,6-trinitrophenyl hapten conjugated to lipopolysaccharide (TNP-LPS), a model antigen. For this, rainbow trout peripheral blood leukocytes were incubated with different doses of ß-glucans or media alone in the presence or absence of TNP-LPS for 48 h. The size, levels of expression of surface MHC II, antigen processing and phagocytic capacities and proliferation of IgM+ B cells were then studied by flow cytometry. The number of IgM-secreting cells in the cultures was also estimated by ELISpot. ß-glucans significantly decreased the levels of surface MHC II expression and the antigen processing capacities of these cells, especially in the presence of TNP-LPS, while they increased their phagocytic activity. On their own, ß-glucans slightly activated the proliferation of IgM+ B cells but reduced that induced by TNP-LPS. In contrast, ß-glucans significantly increased the number of cells secreting IgM in the cultures. This effect of ß-glucans on the IgM-secreting capacity of B cells was also confirmed through a feeding experiment, in which the IgM-secreting capacity of blood leukocytes obtained from fish fed a ß-glucan-supplemented diet for one month was compared to that of leukocytes obtained from fish fed a control diet. Altogether, these findings contribute to increase our knowledge regarding the effects of ß-glucans on fish adaptive responses.

Recalculating the staff required to run a modern assisted reproductive technology laboratory.

STUDY QUESTION: What number of staff is sufficient to perform increasingly complicated processes in today's modern ART laboratories? SUMMARY ANSWER: The adequate number of personnel required for the efficient and safe operation of modern ART laboratories needs to be calculated. WHAT IS KNOWN ALREADY: In today's modern ART laboratories, the amount of time required to perform increasingly complicated processes has more than doubled, with a downward trend in the amount of work an embryologist can do. Different workload unit values have been used to evaluate each workload task and efficiency in a particular ART laboratory, as well as to occasionally compare one laboratory with another. STUDY DESIGN, SIZE, DURATION: Seven senior embryologists working at different IVF centers, three public and four private centers, participated in this multicenter study conducted between 2019 and 2020. We prepared a survey to create a calculator for staff using the average (of three attempts) time spent in every laboratory by each embryologist of the center to perform any ART process. PARTICIPANTS/MATERIALS, SETTING, METHODS: Different laboratory processes and activities related to quality control, time spent and conventional human double witnessing were included in the survey. To calculate the number of processes that each embryologist can perform per year, an embryologist was considered to be having a full-time contract and working 7 or 8 h/day. The times included in the calculation of each task were those corresponding to the 95th percentile. For the calculations, Microsoft® Office Excel® Professional Plus 2019 was used. MAIN RESULTS AND THE ROLE OF CHANCE: The survey showed that the time needed per embryologist to perform the different processes necessary for a classic IVF cycle without time lapse (TL) was 8.11 h, and with TL, it was 10.27 h. The calculated time also considered the time spent in documentation handling, cycle preparation, database management and conventional human double witnessing verification. An ICSI without TL needed 8.55 h, and with TL, it needed 10.71 h. An ICSI-PGT without a TL cycle needed 11.75 h, and with TL, it needed 13.91 h. Furthermore, 1.81 h should be added for every vitrification support needed. The time needed to control more than 200 critical steps, including equipment control and culture parameters, was 30 min per day plus 3.9 min per device to control.The time spent in semen analysis (including documentation handling, cycle preparation and database management) or intrauterine insemination with a partner sperm was 2.7 h. For donor sperm, an additional hour was required for the management involved. The time required to perform a testicular biopsy and cryopreserve the sample was 4 h. Similarly, the time required to perform seminal cryopreservation was 3.7 h. LIMITATIONS, REASONS FOR CAUTION: The study was conducted considering a full-time contract embryologist working 7 or 8 h/day, 5 days a week, with days off according to the Spanish regulations. However, our findings can be adapted to foreign regulations using the developed online calculation platform. WIDER IMPLICATIONS OF THE FINDINGS: A new advanced staff calculator allows any IVF laboratory to estimate the minimum number of embryologists necessary without compromising the security or success of the results. Nevertheless, we recommend a minimum of two qualified embryologists in every laboratory, regardless of the workload. STUDY FUNDING/COMPETING INTEREST(S): This work was funded by the Asociación para el Estudio de la Biología de la Reproducción (ASEBIR). None of the authors has any conflict of interest to declare. TRIAL REGISTRATION NUMBER: N/A.

Mucosal and systemic immune effects of Bacillus subtilis in rainbow trout (Oncorhynchus mykiss).

Bacillus spp. are well known for their probiotic properties. Hence, the long-term feeding of Bacillus spp. strains to different fish species has been proved to confer beneficial effects regarding growth or pathogen resistance, among others. However, whether these strains could function as mucosal adjuvants, up-regulating immune responses after a single administration, has not yet been investigated in fish. Thus, in the current work, we have performed a series of experiments in rainbow trout (Oncorhynchus mykiss) aimed at establishing the potential of two Bacillus subtilis spore-forming strains, designated as ABP1 and ABP2, as oral adjuvants/immunostimulants. As an initial step, we evaluated their transcriptional effects on the rainbow trout intestinal epithelial cell line RTgutGC, and in gut tissue explants incubated ex vivo with the two strains. Their capacity to adhere to RTgutGC cells was also evaluated by flow cytometry. Although both strains had the capacity to modulate the transcription of several genes related to innate and adaptive immune responses, it was the ABP1 strain that led to stronger transcriptional effects, also exerting a higher binding capacity to intestinal epithelial cells. Consequently, we selected this strain to establish its effects on splenic B cells upon in vitro exposure as well as to determine the transcriptional effects exerted in the spleen, kidney, and gut after a single oral administration of the bacteria. Our results showed that B. subtilis ABP1 had the capacity to modulate the proliferation, IgM secreting capacity and MHC II surface expression of splenic B cells. Finally, we confirmed that this strain also induced the transcription of genes involved in inflammation, antimicrobial genes, and genes involved in T cell responses upon a single oral administration. Our results provide valuable information regarding how B. subtilis modulates the immune response of rainbow trout, pointing to the usefulness of the ABP1 strain to design novel oral vaccination strategies for aquaculture.

Atlantic salmon post-smolts adapted for a longer time to seawater develop an effective humoral and cellular immune response against Salmonid alphavirus.

Salmonid alphavirus (SAV) causes pancreas disease (PD) in Atlantic salmon (Salmo salar L.) and disease outbreaks are mainly detected after seawater transfer. The influence of the smoltification process on the immune responses, specifically the adaptive response of Atlantic salmon after SAV infection, is not fully understood. In this study, Atlantic salmon post-smolts were infected by either bath immersion (BI) or intramuscular injection (IM) with SAV subtype 3, 2 weeks (Phase A) or 9 weeks (Phase B) after seawater transfer. The transcript levels of genes related to cellular, humoral and inflammatory responses were evaluated on head kidney samples collected at 3, 7, 14, 21, and 28 days post-infection (dpi). Corresponding negative control groups (CT) were established accordingly. Significant differences were found between both phases and between the IM and BI groups. The anti-inflammatory cytokine IL-10 was up-regulated in Phase A at a higher level than in Phase B. High mRNA levels of the genes RIG-1, SOCS1 and STAT1 were observed in all groups except the BI-B group (BI-Phase B). Moreover, the IM-B group showed a higher regulation of genes related to cellular responses, such as CD40, MHCII, and IL-15, that indicated the activation of a strong cell-mediated immune response. CD40 mRNA levels were elevated one week earlier in the BI-B group than in the BI-A group (BI-Phase A). A significant up-regulation of IgM and IgT genes was seen in both IM groups, but the presence of neutralizing antibodies to SAV was detected only in Phase B fish at 21 and 28 dpi. In addition, we found differences in the basal levels of some of the analysed genes between non-infected control groups of both phases. Findings suggest that Atlantic salmon post-smolts adapted for a longer time to seawater before they come into contact with SAV, developed a stronger humoral and cell-mediated immune response during a SAV infection.

Trace metals and oxidative status in soft tissues of caged mussels (Aulacomya atra) on the North Patagonian coastline.

This study investigated metal accumulation and oxidative effects in mantle, gill and digestive gland of the ribbed mussel Aulacomya atra from the Argentinean North Patagonian coastline. Mussels were transplanted over an 18-month period from a site with low anthropogenic impact to a harbor site with higher seawater concentration of aluminum, chromium, copper, manganese, nickel and zinc. Total trace metal concentration in seawater did not change throughout the 18-month transplant in either site. A. atra bioaccumulated metals in digestive gland, gills and mantle at different levels. Digestive gland had the highest concentration of metals, especially towards the end of the transplant experiment in the harbor area. Mussels transplanted to the harbor site experienced an upregulation in their antioxidant system, which likely explains the lack of oxidative damage to lipids despite higher metal accumulation. These results demonstrate that A. atra selectively accumulates metals from the water column and their prooxidant effects depend on the tissue antioxidant defenses and the exposure time.

Evolution of lymphocytes. Immunoglobulin T of the teleost sea bass (Dicentrarchus labrax): Quantitation of gene expressing and immunoreactive cells.

Immunoglobulin T (IgT) is one of the key effector molecules of jawed vertebrate's adaptive immune system, and in this work we describe the quantitative distribution of IgT-expressing and IgT-producing cells in tissues of the European seabass Dicentrarchus labrax by using mRNA riboprobes and a specific anti-IgT antibody. A polyclonal antiserum (pAb) was prepared by immunizing rabbits with three synthetic peptides deduced from the full length IgT cDNA sequence and located in a surface-exposed CH3 domain of IgT constant region. The obtained antiserum, named RAIgT1, was able to recognize by ELISA immunization antigens and IgT from intestinal mucus and serum. In western blots of head kidney leukocytes lysates the antiserum recognized a 180 kDa polypeptide in non-reducing, and a 75 kDa peptide in reducing conditions. Interestingly, the RAIgT1 pAb crossreacted intensely in western blots with rainbow trout IgT purified from mucus and serum. Antisense mRNA IgT oligonucleotide sequences were employed in in situ hybridization to detect IgT-expressing cells in sections from lymphoid tissues, and positive cells were observed in head kidney, spleen, intestine and gills. By employing RAIgT1 in quantitative immunohistochemistry, the highest number of IgT-producing cells was observed in the gills (9.5 ± 0.7%), followed by intestine (8.4 ± 1.2%), head kidney (6.2 ± 1.4%), and spleen (4.1 ± 0.7%). Interestingly, the number of IgT-B cells showed a regionalization in the intestine, increasing from the proximal to the terminal part. By immunofluorescence and flow cytometry of live leukocytes, the percentages of RAIgT1 stained cells were 34 ± 11% in the intestine, 22 ± 5% in head kidney, 16 ± 7% in spleen, and 9 ± 5% in gills. At the fluorescence microscope, live cells from these tissues showed a typical membrane-associated positivity and a lymphocytic morphology, and no IgT/IgM double positive cells were detected. Immunoreactive cells have been purified from head kidney using magnetic beads, and IgT-enriched cells showed by RT-PCR an enhanced expression of the IgT gene, whereas IgT-depleted cells had an highest expression of IgM and TRß genes. These data describe for the first time a quantitative panel of IgT-expressing and IgT-immunoreactive cells in tissues of a teleost fish species.

Quantitative immunoenzymatic detection of viral encephalopathy and retinopathy virus (betanodavirus) in sea bass Dicentrarchus labrax.

Viral encephalopathy and retinopathy disease caused by betanodavirus, genus of the family Nodaviridae, affects marine, wild and farmed species including sea bass, one of the most important farmed species in Europe. This work describes a reliable and sensitive indirect ELISA assay to detect betanodavirus in biological samples using a polyclonal antiserum (pAb 283) against the 283/I09 virus strain, the most common red-spotted grouper nervous necrosis virus (RGNNV) genotype in the Mediterranean area, and a capture-based ELISA using a monoclonal antibody (mAb 4C3) specific to a common epitope present on the capsid protein. Using adsorbed, purified VERv preparation, the detection limit of indirect ELISA was 2 µg mL(-1) (3 × 10(5) TCID50 per mL), whereas for capture-based ELISA, the sensitivity for the antigen in solution was 17 µg mL(-1) (35 × 10(5) TCID50 per mL). The capture-based ELISA was employed to detect VERv in brain homogenates of in vivo infected sea bass and resulted positive in 22 of 32 samples, some of these with a high viral load estimates (about 1.1 × 10(8)  TCID50 per mL). The ELISA system we propose may be helpful in investigations where coupling of viral content in fish tissues with the presence of circulating VERv-specific IgM is required, or for use in samples where PCR is difficult to perform.

[Risk factors for the deterioration of quality of life in critical trauma patients. Assessment at 6 and 12 months after discharge from the intensive care unit]. / Factores de riesgo de deterioro de calidad de vida en pacientes traumáticos críticos. Valoración a los 6 y 12 meses del alta de la unidad de cuidados intensivos.

OBJECTIVE: To evaluate factors influencing the deterioration of health-related quality of life (HRQoL) in trauma patients admitted to an ICU. DESIGN: A prospective observational study was carried out. SETTING: The combined medical/surgical ICU in a university secondary hospital with 24-hour neurosurgery service. PATIENTS: Trauma patients admitted to the ICU during a two-year period. HRQoL assessment prior to admission to the ICU, and at 6 and 12 months after discharge. MAIN VARIABLES: Demographic variables, type and severity of injury (AIS), severity (APACHE II, ISS, TRISS), length of stay, procedures, mortality and HRQoL according to the SF-36 and EQ-5D. RESULTS: We completed the monitoring of 110 patients that showed significant impairment of their HRQoL in all the dimensions assessed. According to the SF-36, physical role was more deteriorated at 12 months, but the mental component decreased more than the physical component after 6 months. The VAS scale of the EQ-5D decreased to 55 at 6 months (19) and increased to 66 at 12 months (17). In the multiple logistic regression analysis, the variables associated with poorer HRQoL were age > 45 years, TRISS > 10, previous porer quality of life, and serious injuries in the extremities. CONCLUSIONS: Patients showed marked deterioration of their HRQoL at 6 months, followed by overall improvement at 12 months, though without reaching their previous state. The factors that determine poorer quality of life include age, severity, previous HRQoL, and severe injuries in the extremities.

[Multidisciplinary care during peripartum: mother-baby dyad in the center of the relational constellations]. / Prise en charge multidisciplinaire du péripartum: le lien mère-bébé au centre des constellations relationnelles.

Pregnancy and new motherhood may be crisis and vulnerability periods and therefore increase the risk of psychiatric disorders. Liaison psychiatry plays a major role in the first psychiatric evaluation of mothers in order to specify a diagnosis and to initiate a treatment when necessary. This article describes the care of mothers suffering from peripartum psychiatric disorders by the liaison psychiatry in the maternity ward, an outpatient practice, as well as an in-patient care unit where mother and baby can stay together. The multidisciplinary approach and its constellation around the mother-baby dyad are detailed and two clinical cases are reported.

Implementation of a major trauma team. Analysis of activation and care times in patients admitted to the ICU.

OBJECTIVE: To analyze the factors associated with the activation of the severe trauma care team (STAT) in patients admitted to the ICU, to measure its impact on care times, and to analyze the groups of patients according to activation and level of anatomical involvement. DESIGN: Prospective cohort study of severe trauma admitted to the ICU. From June 2017 to May 2019. Risk factors for the activation of the STAT analysed with logistic regression and CART type classification tree. SETTING: Second level hospital ICU. PATIENTS: Patients admitted consecutively. INTERVENTIONS: No. MAIN VARIABLES OF INTEREST: STAT activation. Demographic variables. Injury severity (ISS), intentionality, mechanism, assistance times, evolutionary complications, and mortality. RESULTS: A total of 188 patients were admitted (46.8% of STAT activation), median age of 52 (37-64) years (activated 47 (27-62) vs not activated 55 (42-67) P = 0.023), males 84.0%. No difference in mortality according to activation. The logistic model finds as factors: care (16.6 (2.1-13.2)) and prehospital intubation (4.2 (1.8-9.8)) and severe lower extremity injury (4.4 (1.6-12.3)). Accidental fall (0.2 (0.1-0.6)) makes activation less likely. The CART model selects the type of trauma mechanism and can separate high and low energy trauma. CONCLUSIONS: Factors associated with STAT activation were prehospital care, requiring prior intubation, high-energy mechanisms, and severe lower extremity injuries. Shorter care times if activated without influencing mortality. We must improve activation in older patients with low-energy trauma and without prehospital care.

TB in the elderly: clinical features and outcomes.

BACKGROUND: TB in low-incidence countries is characterised by changes in age distribution towards larger numbers of cases among the elderly.OBJECTIVES: To investigate clinical features and outcomes of TB treatment in older patients and identify predictors of poor outcome.METHODS: Multicentre retrospective study of new TB cases from 53 hospitals included in the registry of the Integrated Tuberculosis Research Programme of the Spanish Society of Pulmonology and Thoracic Surgery (Sociedad Española de Neumología y Cirugía Torácica) between 2006 and 2020.RESULTS: We identified 731 patients aged ≥75 years from a cohort of 7,505 patients with TB. In the elderly, weight loss, disseminated disease and normal X-rays or infiltrates without cavitation were more common. All-cause mortality was 16% (5% of deaths due to TB). The elderly had higher rates of toxicity (6.7%) and hospital admissions (36%). In the multivariate analysis of predictors of TB mortality in ≥75-year-olds, only weight, age and treatment with non-standard regimens remained significant.CONCLUSIONS: TB in older patients needs more attention and remains a challenge because of a lack of specific clinical and radiological features. Standard treatment is effective, although mortality is higher than in young patients. Low weight, non-standard regimens and age are significant predictors of TB mortality.

Impact of the Management and Proportion of Lost to Follow-Up Cases on Cancer Survival Estimates for Small Population-Based Cancer Registries.

BACKGROUND: Estimation of survival requires follow-up of patients from diagnosis until death ensuring complete and good quality data. Many population-based cancer registries in low- and middle-income countries have difficulties linking registry data with regional or national vital statistics, increasing the chances of cases lost to follow-up. The impact of lost to follow-up cases on survival estimates from small population-based cancer registries (<500 cases) has been understudied, and bias could be larger than in larger registries. METHODS: We simulated scenarios based on idealized real data from three population-based cancer registries to assess the impact of loss to follow-up on 1-5-year overall and net survival for stomach, colon, and thyroid cancers-cancer types with very different prognosis. Multiple scenarios with varying of lost to follow-up proportions (1-20%) and sample sizes of (100-500 cases) were carried out. We investigated the impact of excluding versus censoring lost to follow-up cases; punctual and bootstrap confidence intervals for the average bias are presented. RESULTS: Censoring of lost to follow-up cases lead to overestimation of the overall survival, this effect was strongest for cancers with a poor prognosis and increased with follow-up time and higher proportion of lost to follow-up cases; these effects were slightly larger for net survival than overall survival. Excluding cases lost to follow-up did not generate a bias on survival estimates on average, but in individual cases, there were under- and overestimating survival. For gastric, colon, and thyroid cancer, relative bias on 5-year cancer survival with 1% of lost to follow-up varied between 6% and 125%, 2% and 40%, and 0.1% and 1.0%, respectively. CONCLUSION: Estimation of cancer survival from small population-based registries must be interpreted with caution: even small proportions of censoring, or excluding lost to follow-up cases can inflate survival, making it hard to interpret comparison across regions or countries.

Feeding behaviour of Belminus ferroae (Hemiptera: Reduviidae), a predaceous Triatominae colonizing rural houses in Norte de Santander, Colombia.

Belminus ferroae (Hemiptera: Reduviidae) was collected from houses in the localities of San Alberto and Santa Catalina in the municipality of Toledo, Department of Norte de Santander, Colombia. Precipitin tests revealed that the main hosts of B. ferroae are insects belonging to the order Blattodea, whose haemolymph serves as food (89.43%). A low rate of haematophagy was indicated on rodents, dogs and humans (7.32%) and no natural infection with Trypanosoma cruzi was found by either microscopic examination or polymerase chain reaction. The finding of B. ferroae inside houses, feeding primarily on Blattodea with only a minor tendency to feed on vertebrate blood, is indicative of both the evolutionary path from predator to blood-sucker taken by Triatominae and their tendency to adapt from silvatic to domestic habitats.

Effect and associated factors of a clinical pharmacy model in the incidence of medication errors (EACPharModel) in the Hospital Pablo Tobón Uribe: study protocol for a stepped wedge randomized controlled trial (NCT03338725).

BACKGROUND: According to WHO, medication error (ME) is a subject that requires attention at all levels of care to reduce severe and preventable damage related to medication use. Clinical pharmacy practice standards have been proposed around the world so that the pharmacist, as part of a multidisciplinary health team, can help improve patient safety; however, further evidence derived from adequate studies is needed to demonstrate this. This study aims to assess the effect of a clinical pharmacy practice model (CPPM) in preventing MEs associated with the medication use process. METHODS: A prospective, stepped-wedge, cluster-randomized, controlled trial with a duration of 14 months will be performed to compare the effect of a CPPM along with the usual care process of patients in the Pablo Tobón Uribe Hospital (Medellin, Colombia). The study is designed as a cluster-randomized controlled trial, involving five hospital wards (clusters) and 720 patients. Medical wards are allocated to interventions using a stepped-wedge design. Clusters are initially assigned to the control group. After a 2-month observation period, hospital clusters were randomly allocated to the intervention group. Study outcomes will be assessed at baseline and at 2, 4, 6, 8, 10, and 12 months after randomization. The primary outcome will be to assess the effect of a CPPM on the incidence of medication errors associated with the medication use process. Drug-related problems and factors that contribute to the occurrence of MEs will be assessed as secondary outcomes. Statistical analyses will be performed using a mixed model, with the treatment group and time as fixed effects and the clustering structure as a random effect. Statistical analysis will be performed using Pearson chi-square tests and Student's t-tests, and a P value < 0.05 will be considered statistically significant. DISCUSSION: As far as we know, this is the first stepped-wedge, cluster-randomized, controlled trial designed to assess the change of a CPPM on the incidence of medication errors in a hospital in Colombia, and it could generate valuable information about a standardized and patient-centered clinical pharmacy model to improve the safety of inpatient care. TRIAL REGISTRATION: ClinicalTrials.gov, NCT03338725. Registered on 9 November 2017. The first patient was randomized on 2 February 2018. PROTOCOL VERSION: 0010112018JG.

Percutaneous Needle Electrolysis Reverses Neurographic Signs of Nerve Entrapment by Induced Fibrosis in Mice.

Nerve entrapments such as carpal tunnel syndrome are the most common mononeuropathies. The lesional mechanism includes a scarring reaction that causes a vascular compromise. The most effective treatment is surgery, which consists of removing the scarred area, thus reverting the vascular impairment. In the present study, a more conservative therapeutic approach has been undertaken to release the nerve by means of galvanic current (GC) applied with a needle: percutaneous needle electrolysis (PNE). For this purpose, a mouse model of sciatic nerve entrapment has been created using albumin coagulated by glutaraldehyde (albumin 35% and glutaraldehyde 2% volume applied, 10 µl). After two weeks, a fibrous reaction was obtained which entrapped the nerve to the extent of causing atrophy of the leg musculature (14.7%, P < 0.05 compared to the control leg). Ultrasound imaging confirmed that the model's image was compatible with that of nerve entrapment in patients. To quantify the degree of entrapment, nerve conduction recordings were made. The amplitude (peak-to-peak) of the compound muscle action potential (CMAPs) decreased by 32.2% (P < 0.05), and the proximal latency increases by 17.7% (P < 0.05, in both cases). In order to release the sciatic nerve, PNE was applied (1.5 mA for 3 seconds and 3 repetitions; 1.5/3/3) by means of a solid needle in the immediacy of perineural fibrosis before and 5 minutes after the application of GC, and the proximal latency shows a decrease of 16% (P < 0.05). The recovery of CMAPs amplitude was about 48.7% (P < 0.05). Three weeks later, the CMAPs amplitude was almost completely recovered (94.64%). Therefore, with the application of GC by means of a solid needle, the sciatic nerve was definitively released from its fibrous entrapment.

Fluorescent carbon nanoparticles synthesized from bovine serum albumin nanoparticles.

Fluorescent carbon based-nanoparticles are one of the emerging nanomaterials. Their preparation is relatively simple, rapid and inexpensive, and they are less toxic compared with metal and semiconductor nanoparticles. Here, we report a simple and reliable method to prepare water-soluble fluorescent carbon nanoparticles (FC-NPs) from nanoparticles made from a protein, bovine serum albumin. The obtained mean size of our carbon nanoparticles is between 3.8 and 3.4 nm, and they exhibit its maximum fluorescence emission at 424 and 408 nm respectively (with a reasonable QY of 16.5%) due to the presence of functional groups (NH, NH2, COOH and OH) that contain O and N; the presence of these functional groups was confirmed by FTIR and XPS analysis. The photoluminescent decay lifetime was modeled by a two exponential fit which indicates a contribution from both core and surface states. Also, the preliminary results showed that FC-NPs had a good interaction with HeLa and normal oral epithelial cells; nanoparticles were permeable at the cell membrane and went to the cytosol, and even to the nucleus, in less than 30 min, the fluorescence images of our preliminary results did not show any apparent toxic damage in any of the cell lines.

Effects of hypothyroidism on the mesenteric and omental adipose tissue in rats.

To characterize the influence of hypothyroidism on the endocrine activity of mesenteric and omental adipose tissue (MOAT) and the peripheral regulation of energy balance (EB) in rats, we analyzed food intake (FI); basal metabolic rate (BMR); locomotor activity; body weight (BW); serum hormone concentrations and the expression of their receptors in MOAT. We evaluated the morphology and differentiation of adipocytes. Hypothyroidism decreased FI, BMR and BW. The percentage of visceral white adipose tissue (WAT) depots and the morphology of adipocytes were similar to euthyroid rats. Serum leptin and adiponectin expression in MOAT were altered by hypothyroidism. The expression of Perilipin 1, HSL, UCP1 and PRDM16 was significantly lower in MOAT of hypothyroid animals. Hypothyroidism in rats leads to a compensated EB by inducing a white adipocyte dysfunction and a decrease in BW, BMR, FI and adipokine secretions without changing the percentage of WAT depots and the morphology of the MOAT.

Effects of interleukin-1alpha, interleukin-1 receptor antagonist, and neutralizing antibody on proinflammatory cytokine expression by human squamous cell carcinoma lines.

Proinflammatory cytokines interleukin (IL)-1alpha, IL-6, IL-8, and granulocyte macrophage colony-stimulating factor (GM-CSF) have been detected in tumor specimens and primary cell cultures from patients with head and neck squamous cell carcinoma. IL-1alpha has been reported to play an important role in inducing the expression of cytokines IL-6, IL-8, and GM-CSF during inflammation. We examined whether these cytokines are expressed together in five primary and seven established UM-SCC cell lines, and we also examined the effects of IL-1alpha, IL-1 receptor antagonist or neutralizing antibody (Ab) upon expression of this repertoire of proinflammatory cytokines in established UM-SCC lines. Secretion of proinflammatory cytokines IL-1alpha, IL-6, IL-8, and GM-CSF was detected by ELISA in both the primary and established UM-SCC lines. Constitutive expression of specific mRNAs for these cytokines was confirmed in the UM-SCC lines by reverse transcriptase-PCR and Northern blot analysis. Addition of recombinant IL (rIL)-1alpha but not rIL-6 induced a dose-dependent increase in IL-8 and GM-CSF production. IL-1 receptor antagonist (IL-RA) or anti-IL-1 neutralizing Ab could completely inhibit the rIL-1alpha-inducible increase in IL-8 and GM-CSF expression, but the inhibitors had a negligible effect on the constitutive level of production of the cytokines. Transfer and expression of the IL-1alpha gene in a low-cytokine-producing cell line, UM-SCC-38, induced IL-8 and GM-CSF expression, but this expression was also not inhibited by IL-1RA or anti-IL-1 neutralizing Ab. We conclude that IL-1alpha can enhance the expression of cytokines IL-8 and GM-CSF in UM-SCC cell lines but that constitutive expression of these cytokines by UM-SCC is not inhibited by exogenous IL-1RA or neutralizing Ab.