Increase In Secretion Rate Research Articles

Use of Streptococcus salivarius K12 in supporting the mucosal immune function of active young subjects: A randomised double-blind study.

Upper respiratory tract infections (URTI) are the most common illnesses affecting athletes, causing absences from training and competition. Salivary immunoglobulin A (sIgA) is the main immune factor in saliva, and a consistent association between low concentrations of sIgA and an increased incidence of URTIs has been reported. The oral probiotic Streptococcus salivarius K12 has been suggested to have the potential to improve oral diseases and mucosal barrier function. However, the effects of this probiotic on active young subjects performing a high-intensity training (HIT) program have not been investigated. Active young students were randomised into a treated group (S. salivarius K12) and a control (placebo) group and asked to take the product daily for 30 days. After this period, participants performed a graded exercise test and five HIT sessions, all within 3 days. They were also asked to complete the Wisconsin Upper Respiratory Symptom Survey daily to monitor URTI's presence. Before and after the 30 days, and at 0h, 24h and 72h after the last training session, saliva samples were collected to quantify sIgA level, secretion rate, and flow. The effect of S. salivarius K12 intake on these parameters was tested using an ANOVA for repeated measures. Twenty (M = 14, F = 6) young subjects (23.5 ± 2.3 years old) participated in the study. The total accumulated training load (sRPE) in the supplementation period was similar in the two groups (treated: 4345 ± 3441 AU; control: 4969 ± 4165 AU; p > 0.05). Considering both sIgA level and secretion rate, significant time (F(4,15) = 3.38; p = 0.037; F(4,15) = 6.00; p = 0.004) and time×group interactions (F(4,15) = 2.49; p = 0.049; F(4,15) = 5.01; p = 0.009) were reported, with the treated group showing higher sIgA levels at 72h post-exercise and increased secretion rate both at 0h and 72h. The number of URTI episodes was similar in the treated and control groups (χ² = 1.83; p > 0.05). This study demonstrates that relatively short-term S. salivarius K12 supplementation increased sIgA level and secretion in healthy subjects performing a demanding exercise-training programme composed of HIT sessions.

An estimate of extracellular vesicle secretion rates of human blood cells

AbstractExtracellular vesicles (EVs) have been implicated in the intercellular transfer of RNA and proteins through cellular secretion into the extracellular space. In blood plasma, circulating EVs are mainly derived from blood cells; however, factors that control plasma EV abundance are largely unknown. Here, we estimate the EV secretion rates for blood cell types using reported values for cell‐specific plasma EV abundances and their parental cell's ubiquity in healthy humans. While we found that plasma contains on average ∼2 plasma EVs/cell, the cell‐specific EV‐to‐cell ratio spanned four orders of magnitude from 0.13 ± 0.1 erythrocyte‐derived EVs/erythrocyte to (1.9 ± 1.3) × 103 monocyte‐derived EVs/monocyte. The steady‐state plasma EV level was maintained by an estimated plasma EV secretion rate of (1.5 ± 0.4) × 1012 EVs/min. The cell‐specific secretion rate estimates were highest for monocytes (45 ± 21 EVs/cell/min) and lowest for erythrocytes ((3.2 ± 3.0) × 10−3 EVs/cell/min). The estimated basal cell‐specific EV secretion rates were not significantly correlated to the cell's lifespan or size; however, we observed a highly significant correlation to cellular mitochondrial enzyme activities. Together, our analysis indicates that cell‐specific mitochondrial metabolism, for example, via reactive oxygen species, affects plasma EV abundance through increased secretion rates, and the results provide a resource for understanding EV function in human health and disease.

Salivary secretory immunoglobulin A variation between female varsity athletes and nonathletes.

Exercise can lead to changes in immune function. To further investigate this relationship, we examined possible differences in salivary secretory immunoglobulin A (sIgA) secretion rates among female college athletes and nonathlete students. We predicted that athletes would have higher sIgA secretion rates than nonathletes and show greater declines in sIgA during the study period as intensive exercise training continued. We recruited 52 female participants aged 18-22 years (13 swimmers, 20 runners, and 19 nonathletes). We collected two saliva samples from each participant: baseline and a second sample 2 weeks later. Additionally, participants reported minutes of cardiovascular activity performed per week and completed a survey that included a modified Profile of Mood States and the Undergraduate Stress Questionnaire. We found no significant differences in mean salivary sIgA secretion rates between athletes and nonathletes. Salivary sIgA significantly increased between sample collections for both groups, with nonathletes exhibiting a greater increase in sIgA than athletes. Cardiovascular activity was not associated with sIgA secretion rates. Illness status was a significant positive predictor of sIgA secretion rates. Cardiovascular activity did not predict salivary sIgA secretion rates in this sample of female college students, and athletes did not show significantly different sIgA secretion rates than nonathletes. Together, these findings suggest that exercise may not be associated with altered immune function in well-nourished college-aged women. The increase in secretion rates among both athletes and nonathletes suggests that additional factors shared by both groups may have led to increases.

Is sweat testing for cystic fibrosis feasible in patients with down syndrome?

BackgroundRecurrent airway infections are common in patients with Down’s syndrome (DS). Hence, ruling out Cystic Fibrosis (CF) in these patients is often required. In the past, the value of sweat testing – the gold standard to diagnose CF – has been questioned in DS as false positive results have been reported. However, these reports are based on measurements of sweat osmolality or sodium concentrations, not chloride concentrations. This study analyses sweat secretion rate and chloride concentration in sweat samples of patients with DS in comparison to healthy controls.MethodsWe assessed sweat samples in 16 patients with DS and 16 healthy controls regarding sweat secretion rate (SSR) and sweat chloride concentration.ResultsAll measured chloride concentrations were within the normal range. The chloride concentrations were slightly, but not significantly lower in patients with DS (15,54 mmol/l (±4,47)) compared to healthy controls (18,31 mmol/l (±10,12)). While no gender gap in chloride concentration could be found, chloride concentration increased with age in both groups.Insufficient sweat was collected in 2 females with DS (12.5% of the study group) but not in an individual of the control group. A significant lower sweat secretion rate was found in the DS group (27,6 μl/30 min (± 12,18)) compared to the control group (42,7 μl/30 min (± 21,22)). In a sub-analysis, female patients produced significantly less sweat (20,8 ± 10,6 μl/30 min) than male patients with DS (36,4 ± 7,8 μl/30 min), which accounts for the difference between patients and controls. Furthermore, while the sweating secretion rate increased with age in the control group, it did not do so in the DS group. Once again this was due to female patients with DS, who did not show a significant increase of sweat secretion rate with age.ConclusionsSweat chloride concentrations were within the normal range in patients with DS and therefore seem to be a reliable tool for testing for CF in these patients. Interestingly, we found a reduced sweat secretion rate in the DS group. Whether the last one has a functional and clinical counterpart, possibly due to a disturbed thermoregulation in DS patients, requires further investigation.

Incretion of Insulin by Pancreas and Its Inactivation in Women during Long-Term Head-down Tilt Bed Rest

An increase in insular activity in response to hyperglycemia when performing a glucose tolerance test (GTT) during normal physical activity in eight women volunteers was found to be associated with increased blood concentrations of insulin provided by an increase in secretion rate and a decrease in inactivation rate. After 81 days of stay in long-term –6° head-down tilt bed rest (bed rest), flattened glycemic curves (with low hyperglycemic lifting) observed during the GTT, what was indicated high insular activity. An increase in the insular activity was evidencing also by a smaller peak of the blood concentration of glucose from 5 min after intravenous nfusion of 20 mL of 40% glucose solution at 120 minute of the GTT. In bed rest on empty stomach no significant changes in the concentrations of insulin and C-peptide were observed. The dynamics of hormone during the GTT was showed that the appearance of flattened glycemic curves when performing GTT during bed rest was associated with more pronounced, than under the usual conditions, an increase in insulin concentration in response to hyperglycemia. An increase in the blood insulin concentration when performing GTT in bed rest was mainly provided by increased hormone secretion rate in the pancreas, but not by a decrease in its inactivation rate. If you use regular exercises for prevention of hypokinesia under conditions of bed rest, an increase in insulin concentration during GTT was mainly provided by a decrease in its inactivation rate and not by an increase in secretion rate.

Thermogenic and psychogenic recruitment of human eccrine sweat glands: Variations between glabrous and non-glabrous skin surfaces

Human eccrine sweat-gland recruitment and secretion rates were investigated from the glabrous (volar) and non-glabrous hand surfaces during psychogenic (mental arithmetic) and thermogenic stimuli (mild hyperthermia). It was hypothesised that these treatments would activate glands from both skin surfaces, with the non-thermal stimulus increasing secretion rates primarily by recruiting more sweat glands. Ten healthy men participated in two seated, resting trials in temperate conditions (25–26°C). Trials commenced under normothermic conditions during which the first psychogenic stress was applied. That was followed by passive heating (0.5°C mean body temperature elevation) and thermal clamping, with a second cognitive challenge then applied. Sudomotor activity was evaluated from both hands, with colourimetry used to identify activated sweat glands, skin conductance to determine the onset of precursor sweating and ventilated sweat capsules to measure rates of discharged sweating. From glandular activation and sweat rate data, sweat-gland outputs were derived. These psychogenic and thermogenic stimuli activated sweat glands from both the glabrous and non-glabrous skin surfaces, with the former dominating at the glabrous skin and the latter at the non-glabrous surface. Indeed, those stimuli individually accounted for ~90% of the site-specific maximal number of activated sweat glands observed when both stimuli were simultaneously applied. During the normothermic psychological stimulation, sweating from the glabrous surface was elevated via a 185% increase in the number of activated glands within the first 60s. The hypothetical mechanism for this response may involve the serial activation of additional eccrine sweat glands during the progressive evolution of psychogenic sweating.

Evaporimeter and Bubble-Imaging Measures of Sweat Gland Secretion Rates.

Beta-adrenergically-stimulated sweat rates determined by evaporimetry or by sweat bubble imaging are useful for measuring CFTR function because they provide a near-linear readout across almost the full range of CFTR function. They differentiate cystic fibrosis (CF) subjects from CF carriers and carriers from controls. However, evaporimetry, unlike bubble imaging, appears to be unable to detect improved levels of CFTR function in G551D subjects taking the CFTR modulator ivacaftor. Here, we quantify the sensitivity of evaporimetry and bubble imaging methods for assessing low levels of CFTR-dependent sweat rates. To establish sensitivity, we did dose-ranging studies using intradermally injected [cAMP]i–elevating cocktails. We reduced isoproterenol/aminophylline levels while maintaining a high level of atropine to block muscarinic elevation of [Ca2+]i. We stimulated the same sets of glands for both assays and recorded responses for 20 min. In response to a 3-log dilution of the stimulating cocktail (0.1%), bubble responses were detected in 12/12 tests (100%), with 49% ± 3% of glands secreting to produce an aggregate volume of 598 nl across the 12, 20-min tests. This was ~5% of the response to full cocktail. Evaporimetry detected responses in 3/12 (25%) tests with an aggregate secretion volume of 175 nl. After stimulation with a still more dilute cocktail (0.03%), bubble imaging detected 15 ± 13% of glands secreting at a rate ~0.9% of the response to full cocktail, while zero responding was seen with evaporimetry. The bubble imaging method detected secretion down to aggregate rates of <0.2 nl/(cm2·min), or ~1/30th of the average basal transepithelial water loss (TEWL) in the test subject of 4 g/m2·hr or 6.7 nl/(cm2·min). The increased sensitivity of bubble imaging may be required to detect small but physiologically important increases in secretion rates produced by CFTR modulators.

Stress and hypertensive disease: adrenals as a link. Experimental study on hypertensive ISIAH rat strain

ABSTRACTObjective: Association between stress and hypertensive disease is still a matter of debate. Can stress be the cause of hypertensive disease and, if so, what mechanisms are involved? To clarify this question, the Inherited stress-induced arterial hypertensive rat strain (ISIAH rat strain) with a stress related arterial hypertension was developed by selection for the enhanced blood pressure response to 0.5 h restraint stress. The main intention of this work is to confirm that the adrenals are a main link between stress and hypertensive disease. Methods: Hypertensive ISIAH and normotensive WAG rats have been studied. The in vivo secretion rate of corticosterone, aldosterone, 11-Deoxycorticosterone (DOC), and 11-dehydrocorticosterone was measured in anesthetized rats by adrenal vein cannulation. The Dexamethasone/Adrenocorticotropic hormone (DEX/ACTH) test was performed and mRNA expression of Cyp11b1 and Cyp11b2 genes in adrenals was evaluated by real-time PCR. Results: An increased secretion rate of corticosterone and DOC and higher peripheral plasma aldosterone concentration in ISIAH rats were revealed. Response of plasma aldosterone to the surgical stress (adrenal vein cannulation) in the ISIAH rats was significantly higher. The increase of corticosterone and aldosterone in response to ACTH was also higher in hypertensive rats. The basal mRNA expression of both Cyp11b1 and Cyp11b2 genes was increased in the ISIAH rats. The ratio 11-dehydrocorticosterone/corticosterone in ISIAH rats was low which indicates the weakening of 11-beta-Hydroxysteroid dehydrogenase (11-beta-HSD) type 2 converting corticosterone to cortisone. Conclusion: ISIAH rats may serve as a living proof that stress may produce sustained hypertension, and genetically determined enhanced stress responsiveness of corticosterone and, especially, aldosterone may play a crucial role in the mechanism of hypertension development.

Pulse wave myelopathy: An update of an hypothesis highlighting the similarities between syringomyelia and normal pressure hydrocephalus

Most hypotheses trying to explain the pathophysiology of idiopathic syringomyelia involve mechanisms whereby CSF is pumped against a pressure gradient, from the subarachnoid space into the cord parenchyma. On review, these theories have universally failed to explain the disease process. A few papers have suggested that the syrinx fluid may originate from the cord capillary bed itself. However, in these papers, the fluid is said to accumulate due to impaired fluid drainage out of the cord. Again, there is little evidence to substantiate this. This proffered hypothesis looks at the problem from the perspective that syringomyelia and normal pressure hydrocephalus are almost identical in their manifestations but only differ in their site of effect within the neuraxis. It is suggested that the primary trigger for syringomyelia is a reduction in the compliance of the veins draining the spinal cord. This reduces the efficiency of the pulse wave dampening, occurring within the cord parenchyma, increasing arteriolar and capillary pulse pressure. The increased capillary pulse pressure opens the blood–spinal cord barrier due to a direct effect upon the wall integrity and interstitial fluid accumulates due to an increased secretion rate. An increase in arteriolar pulse pressure increases the kinetic energy within the cord parenchyma and this disrupts the cytoarchitecture allowing the fluid to accumulate into small cystic regions in the cord. With time the cystic regions coalesce to form one large cavity which continues to increase in size due to the ongoing interstitial fluid secretion and the hyperdynamic cord vasculature.

Lipid Binding of the Amphipathic Helix Serving as Membrane Anchor of Pestivirus Glycoprotein Erns.

Pestiviruses express a peculiar protein named Erns representing envelope glycoprotein and RNase, which is important for control of the innate immune response and persistent infection. The latter functions are connected with secretion of a certain amount of Erns from the infected cell. Retention/secretion of Erns is most likely controlled by its unusual membrane anchor, a long amphipathic helix attached in plane to the membrane. Here we present results of experiments conducted with a lipid vesicle sedimentation assay able to separate lipid-bound from unbound protein dissolved in the water phase. Using this technique we show that a protein composed of tag sequences and the carboxyterminal 65 residues of Erns binds specifically to membrane vesicles with a clear preference for compositions containing negatively charged lipids. Mutations disturbing the helical folding and/or amphipathic character of the anchor as well as diverse truncations and exchange of amino acids important for intracellular retention of Erns had no or only small effects on the proteins membrane binding. This result contrasts the dramatically increased secretion rates observed for Erns proteins with equivalent mutations within cells. Accordingly, the ratio of secreted versus cell retained Erns is not determined by the lipid affinity of the membrane anchor.

Investigation into the Molecular Mechanisms Underlying Idiopathic Intracranial Hypertension (P2.099)

Objective: To investigate cerebrospinal fluid (CSF) secretion/drainage rates as putative underlying causes of idiopathic intracranial hypertension (IIH). Background: Various causative factors have been postulated as to the increasing incidence of IIH, however, none with any convincing evidence. Identifying biomarkers, already found to be elevated in patients with IIH, including pro-inflammatory cytokines; chemokine (C-C motif) ligand 2 (CCL2), interleukin (IL)-17 (IL-17), IL-6, IL-1β, tumour necrosis factor-α (TNF-α), as well as adipocyte derived hormone leptin and the glucocorticoid hydrocortisone has been used to present an insight into the pathogenesis of IIH. Design/Methods: To assess the effects of the elevated mediators on CSF volume/secretion rates, an in vivo ventriculo-cisternal perfusion was performed. In addition, an in vivo variable rate infusion technique assessed mediator effects on CSF drainage. Results: Hydrocortisone (2.65 μl/min) and TNF-α (2.58 μl/min) significantly increased CSF secretion rate, when compared to controls (1.90μl/min). IL-6 (0.91 μl/min) caused a significant decrease in CSF secretion rate. The volumes of the initial CSF within the cranium following intraperitoneal mediator injection, prior to perfusion of the animal with artificial CSF containing the mediator, were significantly higher for CCL2 (323.6 μl) and IL-17 (327.0 μl) when compared with controls (226.7 μl). CCL2 (55.6 mmH2O.min/μl) and IL-17 (52.7 mmH2O.min/μl) significantly increased pressure and resistance to CSF drainage when compared with controls (28.76mmH2O.min/μl). Conclusions: An impairment within the CSF drainage mechanisms could be postulated due to the significant increase in initial CSF volume; decrease in overall CSF secretion and increased resistance to CSF drainage associated with CCL2 and IL-17. The opposing results of CSF secretion/drainage suggest an altered compensatory mechanism associated with hydrocortisone, TNF-α and IL-6. Study supported by: Prof. Basil Sharrack, Dr. Ignacio A. Romero, Dr. A. Jane Loughlin, Dr. Jane E. Preston. Disclosure: Dr. Alimajstorovic has nothing to disclose.

Mechanisms of hepatic triglyceride accumulation in non-alcoholic fatty liver disease

Non-alcoholic fatty liver disease (NAFLD) is characterized by hepatic lipid accumulation in the absence of excess alcohol intake. NAFLD is the most common chronic liver disease, and ongoing research efforts are focused on understanding the underlying pathobiology of hepatic steatosis with the anticipation that these efforts will identify novel therapeutic targets. Under physiological conditions, the low steady-state triglyceride concentrations in the liver are attributable to a precise balance between acquisition by uptake of non-esterified fatty acids from the plasma and by de novo lipogenesis, versus triglyceride disposal by fatty acid oxidation and by the secretion of triglyceride-rich lipoproteins. In NAFLD patients, insulin resistance leads to hepatic steatosis by multiple mechanisms. Greater uptake rates of plasma non-esterified fatty acids are attributable to increased release from an expanded mass of adipose tissue as a consequence of diminished insulin responsiveness. Hyperinsulinemia promotes the transcriptional upregulation of genes that promote de novo lipogenesis in the liver. Increased hepatic lipid accumulation is not offset by fatty acid oxidation or by increased secretion rates of triglyceride-rich lipoproteins. This review discusses the molecular mechanisms by which hepatic triglyceride homeostasis is achieved under normal conditions, as well as the metabolic alterations that occur in the setting of insulin resistance and contribute to the pathogenesis of NAFLD.

Ocean Acidification Leads to Counterproductive Intestinal Base Loss in the Gulf Toadfish (Opsanus beta)

Oceanic CO(2) has increased from 280 to 380 μatm since preindustrial times and is expected to reach 1,900 μatm by 2300. In addition, regional upwelling zones exhibit levels up to 2,300 μatm, making exploration at future global projected CO(2) levels ecologically relevant today. Recent work has demonstrated that CO(2) exposure as low as 1,000 μatm induces acidosis in toadfish (Opansus beta), leading to metabolic compensation by retention of blood HCO(3) in an effort to defend pH. Since increased serosal HCO(3) translates to increased HCO(3) rates in isolated intestinal tissue, we predicted that blood elevation of HCO(3) and Pco(2) during exposure to 1,900 μatm CO(2) would increase in vivo base secretion rates. Rectal fluid and CaCO(3) excretions were collected from toadfish exposed to 380 (control) and 1,900 μatm CO(2) for 72 h. Fluids were analyzed for pH, osmolality, ionic composition, and total CO(2). Precipitated CaCO(3) was analyzed for titratable alkalinity, Mg(2+), and Ca(2+) content. Fish exposed to 1,900 μatm CO(2) exhibited higher rectal base excretion rates, higher rectal fluid HCO(3) (mmol L(-1)), and lower fluid Cl(-) (mmol L(-1)) than controls, suggesting increased intestinal anion exchange as a result of the compensated respiratory acidosis. This study verifies that imminent projected CO(2) levels expected by the year 2300 lead to greater intestinal HCO(3) loss, a process that acts against compensation for a CO(2)-induced acidosis.

Relationships among salivary immunoglobulin A, lactoferrin and cortisol in basketball players during a basketball season

The aim of this study was to examine the changes and relationships of immune and stress parameters of basketball players during a basketball season. Eight members of National Taichung University basketball team volunteered to participate. Saliva samples were collected at rest and before the start of practice or competition at seven time points during the intense training, competition and recovery period. Salivary immunoglobulin A (sIgA), cortisol, and lactoferrin were measured during training and competition period and compared with those measured at the fourth recovery week. Relationships among immune and stress parameters were evaluated. Compared with those detected at the fourth recovery week, significant decreases in secretion rates and absolute concentrations of sIgA and lactoferrin were observed at times of intense training and competition. In addition, significant increases in secretion rates and absolute concentrations of salivary cortisol were observed during intense training and competition period and the first week of recovery. Moreover, a significant inverse correlation (r=-0.28; P<0.05) that existed between secretion rates of sIgA and cortisol as well as a positive correlation (r=0.32; P<0.05) that existed between secretion rates of sIgA and lactoferrin was measured. Our results demonstrated that the secreted cortisol was induced and the mucosal immunity of the participants was suppressed during the basketball season. The inverse correlation existed between secretion rates of sIgA and cortisol may indicate a possible role of cortisol in the strenuous exercise-induced immunosuppression. Our results also suggest that a delicate balance may exist between mucosal innate and adaptive immune responses.

Is Subclinical Cushing’s Syndrome an Entity or a Statistical Fallout from Diagnostic Testing? Consensus Surrounding the Diagnosis Is Required before Optimal Treatment Can Be Defined

Subclinical hypercortisolism (SH) is a commonly used term to define subtle cortisol hypersecretion but in the absence of symptoms or signs specific of overt hypercortisolism (1, 2). This subtle cortisol excess has been described in up to 20% of patients with an adrenal incidentaloma. With 7% of subjects aged 70 yr harboring such tumors, SH is therefore potentially a common disease (3, 4). When groups of such patients are evaluated, increased body mass index, hypertension, glucose intolerance, hyperlipidemia, and reduced bone mineral density are reported, in keeping with deleterious effects of hypercortisolism. On this background, several studies have now evaluated the effect of laparoscopic adrenalectomy in reversing the biochemical and clinical features of SH with conflicting results (5–7). The latest retrospective study from Chiodini et al. (8) evaluated 108 patients from four hospitals in Milan referred over a 5-yr period. Forty-one of108patientswerediagnosedashavingSH,andofthese, 25 underwent adrenalectomy in a nonrandomized fashion,and16weretreatedconservatively.Thepaperisnovel in that it also presents data on the 67 patients who did not have SH, 30 of whom had surgery on grounds of unacceptable tumor size. They showed that blood pressure, weight, and fasting glucose responded greatest in the treatedSHgroupwhencomparedwithboththeuntreated SH group and the treated non-SH group; conversely, weight, blood pressure, lipids, and glucose seemed to deteriorate in the nontreated SH group over a mean follow-up of 3 yr. The paper has limitations. It is not prospective or randomized and there was clearly a selection bias in that treatedpatientswereyoungerandhadbiggertumors.The absolutevaluesforoutcomemeasuresdonotalwaysalign with the percent numbers of subjects who had steady, increased, or decreased values from which the headline conclusions are drawn. Thus, 32% of treated SH subjects are stated to have a reduction in body weight after treatment, but the raw data show no overall change in body weight in this group (or in fact any of the other three groups after prolonged follow-up). Blood pressure and fasting glucose did fall in the treated SH group but also fell in the non-SH treated group, an interesting observation in its own right but one that suggests an adrenalectomy-induced effect that is independent of cortisol. The greatest area of concern, however, in this paper, and others of its kind, relates to the diagnosis of SH. First, there is a nomenclature issue here. If we agree that SH is more frequently associated with obesity, glucose intolerance,andreducedbonemass,howisthissubclinicalwhen these are established features of Cushing’s syndrome? As we debated when establishing Endocrine Society guidelines for the diagnosis of Cushing’s syndrome (9), the reality is that some of these patients will undoubtedly have mild Cushing’s syndrome, and the difficulty is securing a diagnosis at low levels of cortisol hypersecretion. Here, obesityanddiabetesarepotentialconfounders;activation ofthehypothalamic-pituitary-adrenalaxiswithincreased cortisol secretion is a hallmark of obesity (10), and demonstrating autonomous cortisol hypersecretion vs. increased secretion rate per se is crucial (9). Second, many patients labeled as having SH are likely to be victims of false-positive testing. No single test to

CD40-activated B cells from patients with systemic lupus erythematosus can be modulated by therapeutic immunoglobulins in vitro

Aberrant signaling within and between B and T cells, considered to be central in systemic lupus erythematosus (SLE), could depend on enhanced CD40-CD154 activation. As a result, autoreactive B cells, normally anergic, differentiate and secrete antibodies attacking several normal tissues. Thus restorating B cell homeostasis might help control this disease. In this study, two facets of SLE B cells were investigated, namely their in vitro response to CD40-CD154 and the effect of treatment with human immunoglobulins for intravenous use (IVIg). Blood samples from SLE patients and healthy volunteers were obtained and used to isolate B cells, which were activated through CD40 in the presence or absence of IVIg. The phenotype, proliferation, and differentiation of the SLE B cells were determined and compared with those of control B cells using flow cytometry and standard ELISA. In this model, CD40-activated SLE B cells, as control B cells, proliferated and differentiated and were characterized by the emergence of CD19(lo)CD38(++)CD138(+)CD27(++) cells. IVIg treatment of the CD40-activated SLE B cells resulted in higher differentiation, characterized by increased secretion rates of IgG and IgM, as reported previously for control B cells. Taken as a whole, such accelerated differentiation of CD40-activated B cells suggests that IVIg may participate in re-equilibration of the antibody repertoire by replacing pathological antibodies by de novo harmless antibodies.

Salinity effects on leaf ion composition and salt secretion rate in Avicennia germinans (L.) L.

The effect of salinity on leaf water relations, and rate and ionic composition of leaf secretion was investigated in plants of Avicennia germinans growing under controlled salinity conditions. Increases in salinity from 0 to 940 mol NaCl m-3 reduced the predawn water potential from -0.56 to -4.16 MPa and the solute potential from -2.27 to -4.48 MPa, whereas the pressure potential remained positive in all treatments. Compared to the control, at 940 mol NaCl m-3 Na+ and Cl- concentrations of leaf sap increased 19 and 7 times, respectively. With the addition of salt the main ions accumulated in leaves were Na+ and Cl- whereas plants grown without salt accumulated K+ in leaves at levels similar to those of Na+ with salt treatments. Secretion rates of Na+ and Cl- ions increased up to the highest salinity treatment, amounting to 59 and 46 times respectively, compared to control plants. The increase in secretion rate contributed to counteract the increase of salt concentration in the leaf. In the treatments receiving salts, the Na+/K+ ratio of leaf sap ranged from 4 to 10, whereas that of the secretion ranged from 24 to 56. The ratio of Cl- vs the rest of anions remained around 3 in the leaf sap of salt-treated plants whereas it varied between 96 and 243 in the secretion. With increasing salinity higher secretion rates maintained the internal ion leaf concentration relatively low. The selective character of the secretion contributes to maintain favourable Na+/K+ and Cl-/anion ratios in the leaf cells.

Cell type‐specific regulation of brain‐derived neurotrophic factor in states of allergic inflammation

Brain-derived neurotrophic factor (BDNF) is a molecule influencing neuronal proliferation and differentiation. In states of allergy, it may orchestrate inflammatory changes by linking the immune system with the nervous system. Because the precise regulation of gene transcription in mast cells MCs is not clear, the present studies assessed the gene regulation of BDNF in this inflammatory cell type. Transcriptional expression of BDNF in human skin was studied in isolated cells using RT-PCR. In situ lesional MC BDNF protein expression was analysed by immunohistochemistry and related to the differential staining of MCs and functional effects of BDNF on HaCaT keratinocytes. BDNF mRNA expression was found in isolated human skin MCs, keratinocytes, and fibroblasts. Also, low levels were found in endothelial cells and melanocytes. BDNF protein expression was found in situ in lesional and non-lesional MCs. A significantly decreased expression of BDNF protein was found in atopic dermatitis lesional MCs when compared with control MC expression. Functional in vitro experiments demonstrated that a decrease in BDNF stimulation led to increased secretion rates for stem cell factor and IL-8 in HaCaT keratinocytes. The demonstration of a decreased level of BDNF gene transcription in lesional MCs points to a differential regulation of MC-released neutrotrophins in cutaneous allergic inflammation. Topically administered neurotrophin receptor-modulating compounds should be receptor target specific and not universally acting in diseases such as atopic dermatitis or allergic asthma.

Homologous desensitization and visualization of the tilapia GnRH type 3 receptor

Two types of gonadotropin-releasing hormone (GnRH) receptors were found in the pituitary of tilapia (t), named GnRHR type 3 (tGnRHR3) and GnRHR type 1, according to phylogenetic analysis. tGnRHR3 is highly expressed in the posterior part of the pituitary which contains LH and FSH cells. We characterized tGnRHR3 in terms of both LH release rate and receptor internalization rate in response to continuous exposure to GnRH. Constant exposure of tilapia pituitary fragments to salmon GnRH analog (sGnRHa) resulted in an increased secretion rate for 3 h, followed by a gradual decline, taking 17–19 h, to the basal secretion rate. A chimera between tGnRHR3 and green fluorescent protein (GFP) was created and used to observe the changes in receptor distribution and translocation, activated by agonist with time. The results suggested that the receptor is initially localized at the plasma membrane and upon activation by a homologous ligand (e.g. sGnRHa) undergoes relatively rapid endocytosis. In summary, the present work demonstrates that tGnRHR3 has already undergone endocytosis after 30 min, while desensitization of LH release occurs only after 17–19 h. It is concluded that for tGnRHR3, internalization of the receptor is not exclusively responsible for the desensitization of LH release.

Localization of a Drosophila DRIP-like aquaporin in the malpighian tubules of the house cricket, Acheta domesticus

Malpighian tubules (Mt) are the primary excretory and osmoregulatory organs of insects, capable of rapidly transporting extraordinary volumes of fluid when stimulated by diuretic factors. In the house cricket, Acheta domesticus, the Mt are composed of three morphologically distinct regions (proximal, mid, and distal). Unlike the dipteran Mt, which have both primary and stellate cells, each region of the Acheta Mt consists of a morphologically uniform cell type. The mid and distal regions are both secretory in function and increase secretion rate in response to dibutyryl cAMP (cAMP). Achetakinin-2, while acting synergistically with cAMP on the mid-Mt, inhibits secretion by the distal Mt, and the effects can be reversed by cAMP. Using an antibody to the water-specific Drosophila aquaporin (DRIP), we demonstrated that DRIP-like immunoreactivity was found in both the distal and mid-Mt. The distribution of the aquaporin altered in response to stimulation and was consistent with the secretory data. The regulation of secretion in Acheta Mt is quite different from that of Drosophila, with both cation and anion/water transport occurring in the same cells. This is the first demonstration of the presence of an insect aquaporin, namely DRIP, in the Mt of an order other than the Diptera.