Scenedesmus obliquus can form inducible defensive morphs under grazing threat. Costs and trade-offs of inducible defense are expected to accompany the benefits of defensive morphs, but are hard to detect under nutrient-sufficient experimental conditions. To test the existence of costs associated with inducible defense, we cultured S. obliquus along resource availability gradients in the presence or absence of infochemical cues from Daphnia, and measured the strength of defensive colony formation and fitness characters. Under the lowest phosphorous concentration, the expression of inducible defensive colony resulted in decreased growth rate, which provides direct evidence for physiological costs. Along the gradient reduction of phosphorous concentration or light intensity, inducible defense in S. obliquus showed a decreasing trend. However, the photosynthetic efficiency of S. obliquus was barely affected by its defense responses, suggesting that the negative correlations between resource availability and colony formation of this alga may be due to resource-based trade-offs in the allocation of limited resources. Thus, our results indicated that expression of inducible defense of S. obliquus was impaired under insufficient phosphorus or light. Furthermore, under severe phosphate deficiency, obvious physiological costs of inducible defense could be detected even though defensive colony formation also decreased significantly.The green alga  Phytoplankters, which supply a major part of primary productivity in waters, play an important role in aquatic ecosystems. However, several processes, e.g., hydromechanical dispersion, sedimentation, and consumption by grazers, may cause major loss of phytoplankton biomass. Phytoplankters have to avoid sinking out of euphotic zones to maintain access to sufficient light. Therefore, large size may be resisted among phytoplankton to reduce settling velocityScenedesmus, which forms defensive colonies under predation pressure, is one of the genera that can form induced defensive phenotypes in freshwater5Scenedesmus commonly reproduces asexually through the formation of autospores, in which the mother cell divides into 2–16 daughter cells8Daphnia and Brachionus during active grazing processes may trigger the formation of colonies instead of unicells to resist mortality from grazers6Scenedesmus, many species can respond to infochemicals released by herbivores feeding on focal algal species, e.g. MicrocystisCoelastrumPhaeocystis, which is abundant in the ocean, can form colonies in response to non-species-specific grazing chemical cues4Since phytoplankters share an apparently refugeless openwater habitat with zooplankton and phytoplanktivorous fish, consumption by herbivores is another major factor contributing to phytoplankton loss. To avoid being grazed, many taxa of algae in euphotic water have evolved various chemical or physical defensive strategies, such as toxin production, spine or colony formation, and silica plates34Scenedesmus spp. have been investigated in many studies15Scenedesmus, two main potential indirect costs coupled with morphological defense are identified: reduction of surface-to-volume ratios and enhancement of sinking riskIn terms of energy requirements, defenses that are induced only in the presence of herbivores are considered more efficient than constitutive defenses under variable grazing riskScenedesmus. Colonies not only settle faster but are also less able to resuspend to upper water compared to unicells. For non-motile phytoplankters, prolonged residence in warm upper water with sufficient illumination is a primary requirement. Higher risk of sinking out of euphotic zone increases the loss of population biomass, and decreases the growth rate for light limitation18Scenedesmus.Apart from the relatively small surface area for nutrient and light absorption, increased sedimentation rate of large defensive coenobiums also have negative effect on the population of Scenedesmus182218Scenedesmus. Nevertheless, when alga is cultured in poor conditions, such as nutritional deficiency or light insufficiency, such conditions may be disadvantageous if the cost of maintaining colonies is high. Colony formation of S. acutus has been reported to be a constitutive defense response under phosphorus-sufficient conditions, but an inducible defense response under P-limited conditionsScenedesmus under nutrient-limited conditions caused by competition23Scenedesmus under different phosphorus concentrations and light intensities.Recent research has tried to reveal the cost associated with induced colony formation in Daphnia filtrate stimulated the increase of colony size under all five P concentrations . A three-way ANOVA indicated that the induced colony formation was significantly decreased by decrease in P concentration . In other P concentrations, no statistically significant decrease was observed between control and Daphnia filtrate groups. A two-way ANOVA was conducted to test the effect of Daphnia filtrate and P concentration on growth rate. The result indicated that both factors had significant influences on the growth rate, but no interaction was detected between the two factors  of S. obliquus was not significantly different between the control and Daphnia filtrate groups . A three-way ANOVA revealed significant positive effects of time  and P concentration on Fv/Fm . Consistent with the ratio of Fv/Fm, Daphnia filtrate had no significant influence  on the effective quantum yield of PSII (ΦPSII), but ΦPSII varied significantly with both time  and P concentration . In the two low P concentration treatments (0.05 and 0.2 mg L−1), ΦPSII decreased from day 5. ETRmax showed an increase on day 3. No significant variation was observed between control and Daphnia filtrate groups. Significant effects of time  and P concentration , as well as significant interactions between the two factors , were observed.The photosynthetic efficiency of S. obliquus, light intensity had significant influence on colony formation . Cmax of S. obliquus at 9 μmol photons m−2 s−1 was less than half of Cmax at 72 μmol photons m−2 s−1. However, no significant difference of Cmax was observed under the light intensities within the range of 18–54 μmol photons m−2 s−1. The decreased growth rate of S. obliquus at 9 μmol photons m−2 s−1 indicated growth inhibition by low light intensity  was lower than that in the light intensity experiment. This was possibly due to the higher initial cell density in the P gradient experiment, and the decrease in intracellular phosphorus quota because of pre-treatment of algal cells in non-phosphorus medium.Similar to the effect of P concentration on inducible defense of ormation . The numormation . Unlike r models . One-wayntensity . Resultsnfluence . The groS. obliquus changed with light intensity with or without Daphnia filtrate. Three-way ANOVA indicated that the ratio of Fv/Fm was influenced significantly by time  and light intensity . The ratio of Fv/Fm increased with increase in light intensity. At the highest light intensity, the initial Fv/Fm was high, but decreased with cultural time. Moreover, a time × light intensity interaction  was found. No significant Daphnia filtrate effect was detected . The three factors all affected the ΦPSII of S. obliquus significantly . ΦPSII was negatively affected by the presence of Daphnia filtrate—it decreased significantly on day 3 at 9 μmol photons m−2 s−1, and on day 5 and 7 at 54 and 72 μmol photons m−2 s−1. Significant time × light intensity  and light × Daphnia filtrate  interactions were also observed. For ETRmax, significant time  and light intensity  effects and time × light intensity interaction  were detected. The effect of Daphnia filtrate on ETRmax was not statistically significant .The photosynthetic efficiency of S. obliquus and the effect of resource gradient on this defense response. At all phosphorus concentrations and light intensity treatments, Daphnia filtrate induced the defensive colony formation of by S. obliquus. Consistent with previous studies18Daphnia filtrate and then decreased, possibly because of the increase of cell densities and the degradation of chemical cues. Colony formation in response to Daphnia infochemicals exhibited a decreasing trend with decreasing environmental resource availability  will reduce the intracellular concentrations of Pi and ATP synthesisi deficiency, photosynthesis is inhibited because of decreased ribulose-1,5-bisphosphate pool sizev/Fm and value of ΦPSII, probably suggesting stress on PSII reaction centers under low P concentrations. ETRmax was also decreased under low P concentrations, suggesting that efficiency of the electron transport chain was affected. Although no significant Daphnia filtrate effect was observed on photosynthetic parameters, growth reduction emerged under phosphorous deficiency, specifically under the lowest P concentration, which is a relatively low level in eutrophic lakesS. obliquus.Both light limitation and phosphate deficiency impaired the grazing-induced defense in S. obliquus was similar to that of phosphorous concentration gradient. In this study, the inducible colony formation decreased significantly at a light intensity of 9 μmol photons m−2 s−1 . Resource insufficiency limits the expression of inducible defensive colony formation, thereby suggesting the existence of trade-offs of intercellular resource allocation between defense and growth. Our observation of growth inhibition of defensive algae under low phosphorus condition suggests that the costs may be detected under specific conditions, such as nutrient deficiency. Studying the costs accompanying the benefits of inducible defense is highly important for understanding the mechanism and evolution of inducible defense. Our work provides evidence supporting the existence of costs and trade-offs of inducible defense in S. obliquus.In conclusion, the expression of inducible defensive colony in Scenedesmus obliquus FACHB-416  can distinctly respond to infochemicals from grazers, such as cladoreans Daphnia spp. and rotifers. This alga is a common species used for inducible defense studies. S. obliquus was cultured with a modified algal growth medium BG–11 in a climate-controlled chamber at a constant temperature of 25 ± 0.5 °C with a light–dark period of 14:10 h.The freshwater green alga Scenedesmus exist in the water of herbivorous zooplanktons such as Daphnia, which feeds on Scenedesmus5Daphnia culture was used as a common method to induce colony formation. In our experiments, the grazers Daphnia magna from a laboratory clone was incubated at a density of 300 individuals per liter for 24 h feeding on S. obliquus at a density of 105 cells ml−1. Then, the water was filtered through a 0.1 μm membrane filter  to eliminate bacteria and other impurities. Another suspension of S. obliquus without D. magna was filtered as control water. In preparing both control and D. magna water filtrates, phosphate was first excluded during culture medium preparation, and was added back at different concentrations after feeding for 24 h as outlined below.The chemical cues that can induce anti-grazing colony formation in −1 of P. K2HPO4 was applied as phosphorous resource and KCl was added to compensate for potassium shortage. The measured initial P concentrations in control groups were 0.049 (±0.009), 0.198 (±0.032), 0.515 (±0.029), 1.059 (±0.007), and 5.028 (±0.225) mg L−1, and in Daphnia filtrate groups were 0.050 (±0.004), 0.201 (±0.004), 0.525 (±0.013), 1.033 (±0.034), and 5.039 (±0.153) mg L−1. After the 7-day experiments, P concentrations decreased to 0.021 (±0.005), 0.060 (±0.011), 0.198 (±0.011), 0.719 (±0.007), 4.307 (±0.225) mg L−1 in control groups and 0.019 (±0.005), 0.069 (±0.006), 0.201 (±0.005), 0.723 (±0.050), 4.291(±0.125) in Daphnia filtrate groups. The measured initial nitrogen concentrations in control and Daphnia filtrate groups were 254.25 (±3.70) and 256.29 (±43.97) mg L−1, and it decreased to 226.17 (±34.57), 207.04 (±63.85), 227.19 (±10.06), 227.39 (±13.52), 209.48 (±16.38) mg L−1 in control groups and 236.55 (±7.54), 223.52 (±12.81), 221.69 (±6.54), 224.95 (±8.59), 217.62 (±24.30) mg L−1 in Daphnia filtrate groups. The total phosphorus and nitrogen concentrations were detected by the ammonium molybdate spectrophotometric method and the alkaline potassium persulfate digestion UV spectrophotometric method (China SEPA)−2 s−1 using fluorescent lights on a 14:10 light:dark cycle. Before the experiment, algal cells were starved in non-phosphate conditions for 3 days to eliminate the interference of intracellular phosphate quota. Starved algal cells were then washed with non-phosphate BG–11 medium, centrifuged at a speed of 662 g for 20 min, and placed into 250 ml Erlenmeyer flasks with different phosphorous concentrations. The initial cell density was about 7.0 × 104 cells ml−1.Five phosphorus concentrations and five light intensity levels were established to investigate the potential costs and trade-offs of forming defensive colonies. In the experiment involving different phosphorus (P) concentrations, modified BG–11 culture media with ammonium as nitrogen source were prepared at 0.05, 0.2, 0.5, 1.0, and 5.0 mg L−2 s−1. Algal cultures were accommodated at respective light intensities for 3 days in advance4 cells ml−1. The experimental cultures were also maintained at 25 °C on a 14:10 light:dark cycle. Both experiments had control groups and Daphnia filtrate-treated groups. Daphnia filtrate-treated groups consisted of 200 ml culture systems of S. obliquus at different P levels or light intensities with 10% (v/v) filtered Daphnia water. Control groups contained 10% filtered control water instead of Daphnia water. The measured initial P concentrations of control groups and Daphnia filtrate groups were 5.442 (±0.162) and 5.412 (±0.105) mg L−1. After the 7-day experiments, P concentrations decreased to 5.018 (±0.339), 4.902 (±0.357), 4.595 (±0.160), 4.691 (±0.214), 4.529 (±0.157) mg L−1 in control groups and 4.882 (±0.399), 4.600 (±0.076), 4.736 (±0.193), 4.761 (±0.104), 4.489 (±0.151) in Daphnia filtrate groups. The measured initial N concentrations in control and Daphnia filtrate groups were 254.66 (±25.20) and 259.95 (±35.77) mg L−1, and it decreased to 230.04 (±43.19), 224.14 (±13.56), 211.93 (±10.06), 229.22 (±11.99), 211.31 (±10.32) mg L−1 in control groups and 234.31 (±5.73), 230.24 (±9.04), 223.12 (±7.88), 218.44 (±11.14), 216.81 (±9.54) in Daphnia filtrate groups. All of the experimental cultures were established in 250 ml Erlenmeyer flasks. Erlenmeyer flasks were rotated artificially twice a day to keep the algal cells in suspension.In the experiment involving different light intensities, culture flasks were placed separately in climate-controlled chambers at five levels of light intensity, i.e., 9, 18, 36, 54, and 72 μmol photons mDaphnia filtrate-treated groups were taken at 9:00 a.m. every day under aseptic conditions. Lugol’s iodine solution was added at 2% as preservative after sampling. Two major quantitative traits, namely, population growth and colony size, were measured. Population growth was calculated based on cell density, which was counted using a hemocytometer (0.1 mm deep) on an Olympus microscope . Eighteen 0.1 mmln (S. obliquus cells ml−1) over time. The mean numbers of cells per particle were also calculated from the above counts . The number of cells per particle (C) versus cultural time (t) at all P concentrations and light intensities were fitted by using a Gaussian distribution:Samples (1 ml) of every control and Cmax is the maximum number of cells per particle, t0 is the time to reach Cmax, and a is the full width at half maximum intensity of the curve. To determine the relationship between resource availability and defensive colony formation, the observed number of Cmax was plotted against P concentration and light intensity.where v/Fm, where Fv  = (Fm − F0), and Fm and F0 are the maximal and minimal chlorophyll fluorescence yield of a dark-adapted suspension, respectively. The effective quantum yield of PSII (ΦPSII) was calculated according to the following expression: (F’m − Fs)/F’m, where Fs and F’m are the stable and maximal chlorophyll fluorescence in light-acclimated algal suspensions, respectively. The electron transport rate (ETR) versus irradiance (PAR) curve was plotted for 20 different PARs within the range of 0–2000 μmol protons m−2s−1. The maximal electron transport rate (ETRmax) was determined from a curve after fitting it to the model proposed by Platt et al.Samples (2 ml) of each group were taken every day for the measurement of photosynthetic efficiency with the PHYTO-PAM phytoplankton analyzer . The maximal efficiency of PSII photochemistry was determined as FDaphnia filtrate as the fixed factors. Three-way ANOVA was used to compare the differences and interactions among different environmental resource levels (P or light separately), Daphnia filtrates, and time. All data were analyzed using SigmaPlot 11.0.All values are presented as mean ±SE. Growth rates and numbers of cells per particle were compared by two-way ANOVA with P concentration or light intensity and How to cite this article: Zhu, X. et al. Costs and trade-offs of grazer-induced defenses in Scenedesmus under deficient resource. Sci. Rep.6, 22594; doi: 10.1038/srep22594 (2016).


Inflammatory processes may cause depression in subsets of vulnerable individuals. Inflammation-associated behavioral changes are commonly modelled in rodents by administration of bacterial lipopolysaccharide (LPS). However, the time frame in which immune activation and depressive-like behavior occur is not very clear. In this study, we showed that systemic administration of LPS robustly increased circulating levels of corticosterone, leptin, pro- and anti-inflammatory cytokines, and chemokines. Serum concentrations of most analytes peaked within the first 6 h after LPS injection and returned to baseline values by 24 h. Chemokine levels, however, remained elevated for up to 96 h. Using an optimized sucrose preference test (SPT) we showed that sickness behavior was present from 2 to 24 h. LPS-induced anhedonia, as measured by decreased sucrose preference, lasted up to 96 h. To mimic the human situation, where depression develops after chronic inflammation, rats were preexposed to repeated LPS administration or subchronic restraint stress and subsequently challenged with LPS. While these procedures did not increase the duration of anhedonia, our results do indicate that inflammation may cause depressive symptoms such as anhedonia. Using our SPT protocol, more elaborate rodent models can be developed to study the mechanisms underlying inflammation-associated depression in humans. Major depressive disorder, or depression, is a serious medical illness with a life time prevalence of around 16% . ClinicaBased on these observations, several rodent models of inflammation-associated depression have been developed. One of the most used models involves administration of bacterial lipopolysaccharide (LPS), which is a potent activator of the immune system. Behavioral studies in rodents have shown that systemic LPS injection induces a sickness response, characterized by hypolocomotion, social withdrawal, fatigue, anorexia, and alterations in sleep patterns and cognition . There aIn a previous study we characterized behavioral changes induced by systemic LPS injection in mice . This woL × W × H: 43 × 32 × 18 cm; Tecniplast, Italy) for one week to acclimate prior to experiments. Procedure rooms were maintained at a temperature of 22 ± 2°C and a humidity of 54 ± 2%, with a 12 : 12 h light-dark cycle (lights on at 06:00 a.m. with a 30-minute sunrise and dusk phase). Unless mentioned otherwise, food and water were available ad libitum. All experimental protocols were approved by the Institutional Ethical Committee on Animal Experimentation, in compliance with Belgian law  and conducted in facilities accredited by the Association for the Assessment and Accreditation of Laboratory Animal Care (AAALAC).All studies were conducted in male Sprague-Dawley rats  weighing 200–220 g on arrival. Animals were housed in groups of 4 in plexiglass individually ventilated cages ( Escherichia coli (serotype 055:B5) was purchased from Sigma-Aldrich and freshly dissolved in sterile saline prior to injection.Lipopolysaccharide (LPS) fromn = 12 per group). Just before and at 1 h, 2 h, 6 h, 24 h, 48 h, 72 h, and 96 h after the immune challenge, rats were briefly anesthetized by inhalation of 2% isoflurane, blood was collected from the tail artery, and the rats were returned to their home cage. Serum was obtained by keeping the blood samples in Vacutainer SST II Advance blood tubes  for 30 minutes at room temperature. Then the samples were centrifuged , aliquoted, flash frozen in liquid nitrogen, and stored at −80°C until analysis.To measure the effect of peripheral LPS administration on serum levels of a selection of analytes, rats were injected i.p. with either vehicle or 0.63 mg/kg LPS (γ (IFN-γ), interleukin- (IL-) 1β, IL-6, IL-10, leptin, monocyte chemoattractant protein-1 (MCP-1), macrophage inflammatory protein-1α (MIP-1α), and tumor necrosis factor-α (TNF-α) were simultaneously determined using a rat cytokine/chemokine magnetic bead panel kit from Merck Millipore. This assay is based on Luminex™ technology in which magnetic beads with a distinct emitting fluorescence pattern are coated onto antibodies that specifically capture individual cytokines. All steps in the assay were conducted according to the manufacturer's instructions. A Bio-Plex 200 System (Bio-Rad) was used to measure the fluorescent signal and the data was analyzed using Bio-Plex Manager 5.0 software (Bio-Rad) with five-parameter logistic regression curve fitting. Cytokine levels below detection limit were assigned a value equal to the lowest detectable value of that cytokine. Cytokine values outside of the average + 3 times standard deviation range were considered outliers and were excluded from all calculations. This was the case for less than 2% of all measured cytokines.Serum levels of chemokine (C-X-C motif) ligand 1 (CXCL1), interferon-Serum concentrations of corticosterone were determined using a commercial ELISA kit supplied by Enzo Life Sciences (Product ID 900-097). All procedures were carried out as per manufacturer's instructions.n = 12 per group) were injected i.p. with 0, 0.31, 0.63, or 1.25 mg/kg LPS and tested at 2 h, 6 h, or 24 h after LPS administration. This dose range of LPS was selected based on results from our previous experiments in mice [The open field test (OFT) and forced swim test (FST) setups were custom-made. In these paradigms, groups of naive rats , set up to detect immobility time and distance moved. In this test, exploratory behavior by the animal was used to measure locomotor activity.The FST setup consisted of four independent plexiglass cylinders (diameter 19 cm) which were flushed and filled with water . The four cylinders allowed testing of four rats per session. A video camera with an infrared filter was fixed onto a frame in front of each cylinder and infrared illumination was provided to allow optimal detection and tracking of the rats. Exactly 2 sec after detection of each individual rat, tracking was started and performed using EthoVision 6.1 software . Each FST test consisted of two sessions: a 15 min training session on the day before LPS administration and a 6 min test session at the relevant time after LPS. Immobility time and distance moved (based on center point of gravity of the detected surface) were measured during each session and the duration of immobility was evaluated as a measure of behavioral despair.L × W × H: 35 × 31 × 16 cm; Tecniplast, Italy) fitted with two 250 mL drinking bottles and ad libitum access to food. Each of the drinking bottles contained either filtered tap water or a 1% sucrose solution. The location of each bottle on the cage was randomized every day, with half the animals receiving sucrose on the left and the other half on the right. Prior to LPS administration, rats were familiarized to the sucrose solution by presenting them with water/water (W/W) or water/sucrose (W/S) for 24 h each on 2 consecutive days. Then, the rats were fluid deprived overnight and injected i.p. with 0, 0.31, 0.63, or 1.25 mg/kg LPS (n = 12 per group). At 2 h, 6 h, and 24 h after LPS administration all rats were exposed to preweighed drinking bottles containing W/S. After 1 h the bottles were removed and weighed using Software Wedge for Windows 1.2 (TAL Technologies).Animals were single-housed in individually ventilated cages  on familiarization day 1 (FAM1) and FAM3 or one water- and one 1% sucrose-filled bottle (W/S) on FAM2 and FAM4 (All of the remaining SPT experiments started with a 4-dayand FAM4 . The bot test phase started by weighing and injecting rats with either vehicle or 0.63 mg/kg i.p. LPS (n = 12 per group). Immediately after LPS administration, the rats were put into their home cage and given access to W/S for 4 consecutive days. This experiment was repeated three times using 12 naive animals per treatment group in each trial and the data was pooled prior to analysis so that the final n = 36 per group.To assess the effect of a single peripheral bolus of LPS on anhedonia over time, then = 12 per group). After the familiarization phase, a preexposure phase was introduced in which rats from the 5 days of vehicle groups were injected i.p. with vehicle while rats from the 5 days of LPS groups received a daily i.p. injection of 0.63 mg/kg LPS for 5 consecutive days , 5 days of vehicle + LPS on test day (5 Veh + LPS), 5 days of LPS + vehicle on test day (5 LPS + Veh), and 5 days of LPS + LPS on test day (5 LPS + LPS) (ive days . All ratn = 12 per group), and a manipulation phase was introduced in between the familiarization phase and the test phase  for 5 consecutive days. To control for possible effects of handling stress, rats from the no stress groups were weighed, handled, and put back in their home cage. All rats had ad libitum access to food and water during the manipulation phase. The test phase started 3 days after the manipulation phase by injecting the rats i.p. with either vehicle or 0.63 mg/kg LPS. Immediately after LPS administration, all animals were presented with W/S for 4 consecutive days.To assess the effect of stress on LPS-induced anhedonia, rats were randomized across 4 experimental groups, that is, no stress + vehicle (NS + Veh), no stress + LPS (NS + LPS), stress + vehicle (S + Veh), and stress + LPS (S + LPS) . Total fluid intake was taken as the total change in volume from both bottles combined, while sucrose preference was calculated as a percentage of consumed sucrose solution of the total fluid intake. A total fluid intake greater than the mean + two times standard deviation was considered to be an invalid measure that probably resulted from leaking bottles. Invalid measures were replaced by the mean of all the bottles either on the relevant side (for W/W) or for either water or sucrose (for W/S). This happened for less than 2% of bottle measurements in any given experiment. Total volume intake was evaluated as a primary measure for sickness behavior , while sucrose preference was used as a measure of anhedonia. Change in body weight was calculated by subtracting the weight at a given time point from the weight at the start of the experiment. These time points are specified for each experiment in the Results.ε) was used in case of repeated measures analysis to correct for potential violation of the sphericity assumption [F-ratio is evaluated with the new degrees of freedom, resulting in a more conservative statistical test. To account for the skewness of the data distribution, concentrations of serum analytes were log-transformed prior to analysis. Significance was accepted for the ANOVAs and rmANOVAs when p < 0.05. When appropriate, post hoc comparisons were made by using an independent samples t-test with a Bonferroni-corrected p value. For consistency between the analysis and the visualization of serum analyte concentrations, the group means and its standard error of the mean (SEM) were back-transformed and visually presented on a logarithmic scale. All other data are expressed as mean ± SEM on a linear scale.Data were analyzed using SPSS Statistics software . Analysis of variance (ANOVA) or repeated measure ANOVA (rmANOVA) was performed to assess the statistical significance of differences across treatment groups. A Greenhouse-Geisser correction epsilon  = 11.1, p < 0.001) and time point  = 15.6, p < 0.001) for total distance travelled. Post hoc analysis indicated that systemic LPS administration reduced locomotor activity in a dose-dependent manner at 2 h  = 12.3, p < 0.001, ε = 0.98). Post hoc analysis indicated that LPS reduced total volume intake at 6 h and 24 h to a similar extent at all doses  = 11.2, p < 0.001, ε = 0.47), CXCL1  = 56.7, p < 0.001, ε = 0.30), IFN-γ  = 39.8, p < 0.001, ε = 0.29), IL-1β  = 14.9, p < 0.001, ε = 0.28), IL-6  = 76.6, p < 0.001, ε = 0.19), IL-10  = 35.1, p < 0.001, ε = 0.22), leptin  = 6.5, p < 0.001, ε = 0.53), MCP-1  = 288.4, p < 0.001, ε = 0.38), MIP-1α  = 51.1, p < 0.001, ε = 0.33), and TNF-α  = 68.2, p < 0.001, ε = 0.23). Post hoc analysis showed that serum levels of corticosterone were elevated at 2 h, 6 h, and 24 h and fell below control values at 48 h after LPS administration  = 563.2, p < 0.001, ε = 0.69) and LPS assignment  = 5.1, p < 0.05, ε = 0.69) for change in body weight during the familiarization phase. Post hoc analysis indicated that rats in the LPS group had a statistically significant lower change in body weight at familiarization day 1 (FAM1) as compared to animals in the vehicle group . This indicates that the caloric value of sucrose did not influence the change in body weight.Based on the results from the first SPT study  and the  familiarization phase, there was a flavor × repeat interaction  = 8.9, p < 0.01) but no effect of LPS assignment. Post hoc analysis indicated that the total volume intake increased substantially on days that rats were exposed to W/S when compared to W/W days. This increase was slightly reduced upon retesting   = 86.2, p < 0.001, ε = 0.83) for change in body weight during the test phase. Post hoc analysis showed that systemic LPS injection reduced weight during the first 2 days after injection (D1 and D2) and that this weight decrease remained statistically significant throughout the test phase  = 50.0, p < 0.001, ε = 0.87). In the first 24 h after administration (D1), LPS reduced total volume intake to less than one-third of the normal daily water intake, suggesting suppression of drinking as a consequence of sickness  = 2.8, p < 0.05, ε = 0.91) was also found for sucrose preference during the test phase. Post hoc analysis revealed that systemic LPS administration reduced sucrose preference close to chance level  on D1 (A time × LPS interaction (preexposure phase) and measuring sucrose preference after an acute LPS injection 3 days later (test phase).The duration of inflammatory processes associated with depression is thought to be chronic rather than acute. In this experiment, the anhedonic response to a more prolonged immune challenge was investigated by first injecting rats with LPS on 5 consecutive days . To evaluate the weight change after a subsequent acute LPS injection, weight measures during the test phase were subtracted from the weight at the start of the test phase. rmANOVA revealed that there was a time × preexposure × LPS interaction  = 13.9, p < 0.001, ε = 0.76) for change in body weight in the test phase. Post hoc analysis indicated that all rats lost weight after receiving the acute LPS challenge  = 24.4, p < 0.001, ε = 0.93) but not of time or preexposure. Rats that were injected with LPS during the test phase had a reduced sucrose preference when compared to vehicle-injected animals  and subsequently injected systemically with LPS three days later (test phase).Stress, a known risk factor for depression, influences immunological responses. To test whether stress impacts on the anhedonic response to an immune challenge, rats were first exposed to 1 h of restraint stress per day for 5 consecutive days  = 145.6, p < 0.001, ε = 0.69) for change in body weight during the manipulation phase. Post hoc analysis indicated that stressed rats continuously lost weight from the first stress session until the last, while nonstressed rats grew steadily during the manipulation phase . Weight changes induced by a subsequent acute LPS challenge were determined by subtracting weight measures from the rats' weight at the beginning of the test phase. There was a time × LPS  = 23.9, p < 0.001, ε = 0.63) and a stress × LPS  = 16.0, p < 0.001, ε = 0.63) interaction for change in body weight during the test phase. Post hoc analysis showed that LPS decreased weight in stressed and nonstressed rats  = 4.5, p < 0.01, ε = 0.75). On the first 2 days after administration, stressed and nonstressed rats that were injected with LPS drank significantly less than animals that received vehicle  = 59.0, p < 0.001, ε = 0.85), but no main effect of time or stress for sucrose preference during the test phase. LPS-treated rats had a lower sucrose preference than vehicle-injected animals, but the lack of main effects of time and stress did not allow further post hoc analysis  had returned to control levels at 24 h, despite the fact that these animals still drank much less than vehicle-treated rats. Rats injected with higher doses of LPS, in contrast, showed reduced total volume intake and decreased sucrose preference at this time. This may indicate that, at a dose of 0.63 mg/kg (and higher), LPS causes more pronounced anhedonia and therefore this particular dose was selected for further experimentation.α, IL-1β, and IL-6 [β and IL-6 in LPS-challenged rats [α remained elevated up to 96 h after treatment. These chemokines play an important role in leukocyte migration and activation, and their serum profile suggests that the immunological response to systemically administered LPS lasted for several days. Future studies should reveal whether levels of circulating chemokines could be used as a biomarker for inflammation-associated depressive symptoms such as anhedonia.While the immunological response to LPS in mice and its relationship to behavioral changes are well documented, this is not the case for rats. Currently available literature on the effect of systemic LPS administration on circulating levels of inflammatory mediators in the rat is limited by the number of analytes measured and/or time points used. We have previously shown that in the mouse LPS administration leads to a rapid release of proinflammatory cytokines such as TNF-and IL-6 . The preged rats . In our ged rats , is alsoged rats . In lineged rats , 24, we β, IL-6, and TNF-α) will induce depression. Nevertheless, these cytokines may cause depressive symptoms such as anhedonia and thus add to the understanding of the pathological mechanisms of depression.Activation of the immune system is known to deregulate the HPA axis, a physiological finding which is frequently observed in depression . In agreIn our extended SPT study we showed that a single bolus of LPS reduces fluid intake in the first 2 days after administration. More specifically, on the first day after administration, LPS decreased total volume intake to approximately one-third of the normal daily intake . This suppressed drinking is a clear indication of sickness and precludes interpreting reduced sucrose preference as a sign of anhedonia. On the second day, however, LPS-injected rats also drank less than vehicle-treated controls but their total volume intake did reach normal daily intake levels. Therefore, the reduced sucrose preference seen on this day can be interpreted as an anhedonic response to the LPS challenge. This became even clearer on the third day where there was no difference in total volume intake between treatment groups, but still a significant reduction in sucrose preference in LPS-injected rats. The fact that rats treated with LPS started to gain weight at day three further indicates that sickness had dissipated at this point. In line with previously reported rat data , 29, LPSIn our model of acute systemic LPS administration, anhedonia was present in the absence of apparent sickness from 2 to 3 days after injection. Depressive episodes in humans, however, can last up to several months . MoreoveStress is a major risk factor for the development of depression . In a se familiarization phase and test phase. It may be possible that this extra week of individual housing and handling confounded measures of sucrose preference after a subsequent systemic LPS injection.In the experiments where we tested the effect of LPS preexposure or stress on the anhedonia response to a subsequent LPS administration, LPS-injected rats showed reduced sucrose preference across the test phase. These findings are not in line with the acute LPS experiment where sucrose preference was only reduced at days 2 and 3 but had recovered at day 4. This discrepancy can potentially be explained by the fact that in our more elaborate experimental protocols a third phase was introduced between theThis study provides a systematic analysis of the time course of cytokine release and behavioral changes following peripheral LPS administration in rats. We report a SPT protocol that includes measurements of total volume intake, sucrose preference, and body weight and demonstrated that, by assessing these measurements and their interaction, this SPT protocol provides a way of separating LPS-induced anhedonia from sickness. This anhedonic response to LPS is robust but only lasts for 2 days. Therefore, caution is needed when studying the mechanisms underlying inflammation-associated depression using a single LPS injection in rats. To model the chronic nature of depression in humans more carefully, our SPT protocol was used to test whether preexposure to repeated LPS administration or subchronic stress influences the anhedonic response to a subsequent LPS challenge. While these procedures did not affect the time course of anhedonia, our results provide useful insights into the behavioral consequences of peripheral immune activation using LPS and may contribute to the development of more elaborate rodent models of inflammation-associated depression.


Helicobacter cinaedi causes bacteremia without characteristic clinical symptoms and is firstly isolated from human immunodeficiency virus (HIV)-positive homosexual men.H. cinaedi bacteremia. Some cases of H. cinaedi bacteremia may require long-term administration of multiple antibiotics prior to the resolution of infection.Here we describe, for the first time case report, two female patients with diabetes who had H. cinaedi in patients with diabetes presenting with bacteremia, especially in patients with poor glycemic control.Therefore, these cases indicate that it is important to consider  Helicobacter cinaedi is a Gram-negative enterohepatic bacterium of the Helicobacteraceae family that was first identified in 1985 among immunocompromised European and American men with human immunodeficiency virus (HIV) infections  was continued. On day 18 after the initial admission, the third blood culture was taken, then she was discharged a second time. Later, we confirmed that her third blood culture was negative in an outpatient clinic and prescribed completion of the antibiotic regimen.Case 1: A 76-year-old female with a history of slowly progressive type 1 diabetes, treated for 30 years. She was being treated for rheumatoid arthritis  level of 7.8% while on a regimen of 50% insulin lispro protamine suspension and 50% insulin lispro injection  at 6 U in the morning and 4 U in the evening, as well as miglitol (10 mg/day) and mitiglinide (450 mg/day). She presented to the emergency unit of our hospital with a fever of 39°C, headache, erythema on the trunk, and joint pain. On admission, her body temperature was 40.2°C, pulse rate 134 beats/min, blood pressure 150/70 mmHg, height 145 cm, and body weight 38 kg. Routine laboratory testing showed an elevated leukocyte count of 11.8 × 103 cells/μL and a C-reactive protein level of 4.49 mg/dL. Serological tests for syphilis and HIV were negative. Soon after starting antibiotic treatment, her fever subsided. Five days later, redness and swelling disappeared; therefore, we changed treatment to cefdinir at 900 mg/day. She was discharged on day 8. The following day, we confirmed H. cinaedi from the blood culture taken on day 1; nevertheless, we decided to adopt a wait-and-see approach because no exacerbation of disease was observed since her initial admission.Case 2: A 47-year-old female diagnosed with type 2 diabetes 2 years before, and treatment started immediately thereafter. However, after a year, she stopped taking medications on her own decision. She presented to emergency department of our hospital with a fever of 38°C and bilateral lower leg pain for three days. She was hospitalized with a diagnosis of lower right leg cellulitis, and we began cefazolin administration at 3 g/day. On admission, her diabetic control was poor ; in addition, her body temperature was 39.2°C, pulse rate 106 beats/min, blood pressure 137/71 mmHg, height 166 cm, and body weight 61 kg. Routine laboratory testing showed an elevated leukocyte count of 13.8 × 10H. cinaedi is a rare infection primarily detected among HIV-positive patients in western countries. In Japan, most patients with H. cinaedi infection were HIV-negative, but immunocompromised (Araoka et al. H. cinaedi settles in the digestive tracts of humans, monkeys, dogs, and cats. Some cases of hospital-acquired outbreaks have also been reported (Rimbara et al. H. cinaedi infection. However, there is no consensus on characteristic clinical symptoms of H. cinaedi infection and H. cinaedi is a fastidious organism, thereby making it difficult to clinically diagnosis.H. cinaedi infections, even though this organism is reportedly sensitive to many antibiotics. In general, cephalosporins, carbapenems, tetracyclines, and aminoglycosides were found to be effective; however, various H. cinaedi isolates in Japan have shown increased resistance to penicillins and resistance to macrolides and quinolones (Kawamura et al. H. cinaedi bacteremia may require long-term administration of multiple antibiotics prior to the resolution of infection.The average separation and processing period of a blood culture to detect H. cinaedi is 5 days (Araoka et al. H. cinaedi bacteremia. Because H. cinaedi develops no characteristic clinical symptoms, it is important to consider H. cinaedi infection, especially in patients with poorly controlled diabetes who experience a relapse of infection after curative antibiotic treatment.In conclusion, two patients with poorly controlled diabetes presented with Informed consent was obtained from all patients for being included in the study and for publication of this Case report.


A vast literature has associated height with numerous factors, including biological, psychological, socioeconomic, anthropologic, genetic, environmental, and ecologic, among others. The aim of this study is to examine, among U.S. women, height factors focusing on health, income, education, occupation, social activities, religiosity and subjective well-being.Data are from the Women's Health Initiative (WHI) Observational Study. Participants are 93,676 relatively healthy women ages 49–79; 83% of whom are White, 17% Non-White. Statistical analyses included descriptive statistics, chi-square and multivariable covariance analyses.The mean height of the total sample is 63.67 inches. White women are significantly taller than Non-White women, mean heights 63.68 vs. 63.63 inches (p = 0.0333).ocial behavior, i.e. attendance at clubs/lodges/groups. Women who reported attendance ‘once a week or more often’ were taller than those who reported ‘none’ and ‘once to 3 times a month’. Means in inches are respectively for: White women–63.73 vs. 63.67 and 63.73 vs. 63.67, p = 0.0027. p = 0.0298; Non-White women: 63.77 vs. 63.61 and 63.77 vs. 63.60, p = 0.0050, P = 0.0094. In both White and Non-White women, income, education and subjective well-being were not associated with height.Among both Non-White and White women height is associated with sHowever, other factors differed by race/ethnicity. Taller White women hold or have held managerial/professional jobs–yes vs. no–63.70 vs. 63.66 inches; P = 0.036; and given ‘a little’ strength and comfort from religion’ compared to ‘none’ and ‘a great deal’, 63.73 vs. 63.66 P = 0.0418 and 63.73 vs. 63.67, P = 0.0130. Taller Non-White women had better health—excellent or very good vs. good, fair or poor–63.70 vs. 63.59, P = 0.0116.Further research in diverse populations is suggested by the new findings: being taller is associated with social activities –frequent attendance clubs/lodges/groups”, and with ‘a little’ vs. ‘none’ or ‘great deal’ of strength and comfort from religion. Height has been a subject of interest, discussion and analyses as early as biblical times. For example, “In the first book of Samuel we read the account of Saul being selected king. While Saul's qualifications for the job were not described in any detail, there is one attribute specifically mentioned: he was tall.” The body of literature on height is global, vast and increasing The general conclusion from the literature cited is: Taller is associated with favorable early environment, nutrition, medical conditions, health, income and education in both men and women. However, there are exceptions: i) the significant association of height and income were not found My paper is data from the WHI Baseline Data Set of 10/16/2003, Women's Health Initiative Observational Study, provided by the National Heart, Lung and Blood Institute; the data set was converted to a SAS file in 2013. This study examines the association between height and some of the factors cited in the literature such as demographics—age, gender, ethnicity, income, education, occupation–health, social, subjective well-being, among relatively healthy women, 49–79 years of age, who participated in the Women's Health Initiative's Observational Study (WHI OS). Its main purpose is to assess a wide variety of important clinical and public health issues. Enrollment was conducted at 40 centers throughout the US. The justification for the WHI study is: “There is a general recognition that few older women have been studied longitudinally and that major questions about prediction of chronic disease in postmenopausal women remain.” “Participants in the observational study were women aged 49–79 , who were ineligible or unwilling to participate in the clinical trial component or were recruited through a direct invitation for screening into the observational study.” “Many potential participants in the clinical trial component of the study were already undertaking a low fat diet or were using hormone replacement therapy and therefore were excluded or declined to participate clinical trial component. These participants were then enrolled into the observational study. Previous research has demonstrated that at the time of WHI enrollment, women undertaking hormone replacement therapy and/or low fat diets generally had healthier lifestyles than those not possessing these behaviors. The effect of the selection process was that women enrolled in the observational study tended to have healthier lifestyles compared to those enrolled in the clinical trial.” The data set consists of 2022 variables including demographics, eligibility for selection, personal information, medical history, reproductive history, family history, personal habits, thoughts and feeling, and other areas. Participants are 93,676 women—83%  White, 17% Non-White– 8% Black , 4% Hispanic ; the remaining 5% Asian/Pacific Islander, American Indian, and subjects of unknown race/ethnicity. Other demographic variables are age, employment, region of country, employment. Measurements and definitions of height, income, wages as well as other variables may vary in the vast literature and research conducted by economists, social scientists, psychologists, epidemiologists and others. Therefore, definitions in the WHI OS Data Set questionnaire for the major variables analyzed are shown as follows:Height, in inches at age 18 or tallest adult height.Income “total family income (before taxes) from all sources within your household in the last year” Income is coded in 9 categories: 1) less than $10,000 (4.5%), 2) $10,000–19,999 (11.7%), 3) $20,000–34,999 (23.3%), 4) $35,000–49,999 (20.1%), 5) $50,000–74,999 (20.2%), 6) $75,000–99,999 , 7) $100, 000–149,999 (6.8%), 8) $150,000 or more (3.9%); and 9) “Don't know” (3%) and a category, missing (4%). The mode is in the $20,000–34,000 category, the median in the $35,000–49,999 category, interpolated median about $43,000. The eight categories, excluding missing and “Don't know” were condensed to 5—1) less than $20,000 (16.16%), 2) $20,000–34,999 (23.31%), 3) $35,000–74,999 (40.24%), 4) $75,0000–99,999 (9.43%), 5) $100,000 or more (10.86%).Education: 1) Didn't go to school (.09%), 2) Grade school (1–4 years) (.38%), 3) Grade school (5–8 years) (1.20%) 4) Some high school (9–11 years) (3.51%), 5) High school diploma or GED (16.15%). 6) Vocational or Training School (9.74%), 7) Some college or Associate Degree (26.49%), 8) College graduate or Baccalaureate Degree (11.39%). 9) Some Postgraduate or professional (11.76%), 10) Master's degree (15.73%), 11) Doctoral Degree  (2.76%), Missing (0.79%). Condensed into 3 categories: 1) less than high school (22.12%). 2) high school to some college (47.63%) 3) college graduate or more (30.36%).General health—“In general, would you say your health is—on a five point scale: 1) excellent', 17.7%, 2) very good, 40.2%, 3) good, 31.7%, 4) fair, 8.8%, 5) poor, 0.9%), ‘missing’ 0.7%.”“Likelihood of Depression”—scaled from 0 to 100—higher more likelihood. Likelihood of depression, a highly skewed continuous variable was dichotomized at less than or equal to the median (0.0073)/greater than the median.“Religion gives strength and comfort”—three categories–none 12.5%, a little 24.0%, a great deal 63.0%, missing, 0.5%.“Attend clubs, lodges, etc.”—6 categories—1) not at all in the past month, 43.9%; 2) once in the past month; 3) 2 or 3 times in the past month; 4) once a week 8.1%; 5} 2 or 6 times a week 5.6%; 6) every day 0.1%; missing 1.4%; condensed—none (43.89%), monthly (40.91%), weekly or more (13.84%).Main job—present job or past job held the longest. Defined as “Managerial, professional specialty  Missing, 4.7%” No–54.02%, Yes—41.23%.Pain– Quality of life subscale on pain. PAIN ranges from 0 to 100 with a higher score indicating a more favorable health state. From the Rand 36-Item Health Survey (SF-36).Satisfied with quality of life, analogous to Cantril's ladder, 0-Satisfied to 10-Dissatisfied.Rate quality of life, analogous to Cantril's ladder, 0-worst, 10-Best. ‘Happy’: During the past four weeks ‘Have you been happy’. Six point scale 1 = All, 2 = Most, 3 = A good bit, 4 = Some, 5 = A little bit, 6 = None of the time. (From 36/37). This scale was reversed: All = 6, Most = 5, Good Bit = 4, Some = 3, Little = 2, None = 1.‘Emotional well-being’, ranging from 0 to 100 with a higher score indicating a more favorable health state. The source of the scale is the Rand 36-Item Health Survey (SF-36). Computed from Form 36/37, questions 76, 77, 78, 80, and 82. Source: Rand 36-Item Health Survey (SF-36). Quality of life subscale on emotional well-being ranges from 0 to 100 with a higher score indicating a more favorable health state.‘Social support’ is the sum of nine components. Scores range from 9 to 45, higher scores more support. The 9 components, each ranging from 1) None, 2) A little, 3) Some, 4) most, 5) All–of the time, are: Someone - a) ‘to listen when need to talk’, b) ‘to give good advice’; c) ‘who can take you to the doctor’, d) ‘to have a good time with’, e) ‘to help understand a problem when you need it’, f) ‘to help with daily chores if you are sick’, g) ‘to share your private worries’, h) ‘to do something fun with’, i) ‘to love you and make you feel wanted’.Descriptive statistics (means and standard deviations), chi-square analyses for categorical data, linear regression and multivariable analyses of covariance (GLM) were carried out. Multivariable GLM analyses yielded means, standard errors, and p-values controlling for covariates, and pair-wise p-values by class.Descriptive data from univariate analyses are in Univariate and multivariable covariance analyses for height as the outcome were carried out for the 93,676 participants into three groups a) all, b) Non-White and c) White women. Univariate means for height by demographic, behavioral and subjective well-being variables are in Multivariable analyses included height and seven covariates. Income and education as predictors of subjective well-being, club attendance and religion revealed both congruencies and differences among Non-White and White women. Among White women, income and the subjective well-being variables—happiness, emotional well-being, happiness, satisfaction with life, quality of life and social support—and general health were significantly associated. These variables were also associated with education, with the exception of satisfaction with life. In contrast, Non-White women's subjective well-being variables—emotional well-being, happiness, and satisfaction with life—were not associated with income except for quality of life, P = 0.0095 and social support, P = 0.0007. Associations with education were significant for variables: happiness, emotional well-being and quality of life; satisfaction with life, but not significant for social support.  An additIn sum, new findings from this study of US women, 49–79, are: a) taller Non-White and White women engaged in more frequently in social activities, e.g., such as club attendance; b) taller White women had reported significantly more ‘a little’ strength and comfort from religion compared to ‘none’ and compared to ‘a great deal’. Other major findings are: c) taller Non-White and Whites did not have higher incomes or more education; d) taller White women with present or past managerial/professional jobs; e) taller Non-White women had better general health.A vast and global literature examines the relation of height with numerous factors, including, but not limited to psychological, social, economic, anthropologic, genetic, gender, environmental, ecologic, behavioral, nutritional, infection and other constructs. This study examined data from relatively healthy women ages 49–79, from a range of race/ethnic groups—dichotomized Non-white 17% and White 83% of the sample of 93,676 women. It focused on height and variables including income, education, general health, social activities, and subjective well-being. Two major findings emerge: 1) taller Non-White and White women engaged social activities, viz. attended clubs/lodge/groups, more frequently than those who did not attend or attended less frequently. Attendance at clubs is one among a variety of social activities. Notably, this finding is in accord with Persico et al. 2) Strength and comfort from religion was associated not only with height, but also with subjective well-being, general health, income and education. —4, 7–8. Occupation and height of men and women have been examined by many investigators However, height was not significantly associated with income nor with and education among both Non-white and White. This is in contrast to findings of Deaton and Arora, who analyzed the Analysis of the Gallup-Healthways Well-Being Index daily poll of the US population A new area examined in this study is religiosity as measured as ‘strength and comfort from religion’ classified as ‘none’, ‘a little’ and ‘a great deal’. Overall results are the percentage of women reporting—12% ‘none’, 24% ‘a little’ and 63% ‘a great deal’, and 0.5% missing data. Analyses of this construct, both as a covariate and as a outcome, (to my knowledge has not examined in the literature on height), was related to height, as well as health, subjective well-being, income and education  and 3, AFurther research, suggested by my findings, on height and other factors are the following:Occupation–indicated by the finding that taller White women had managerial/professional jobs presently or in the past. In the WHI data ‘managerial/professional job’ covers a range of occupations'. It is defined as “Managerial, professional specialty ”. To understand better the association of height and the components of ‘managerial/professional specialty need more detailed classifications.Social activities—here denoted by attendance at clubs/lodges/groups—a construct significantly associated with height among Non-White and White. What constitutes social activities and how to measure them needs further work.‘Strength and comfort from religion’, and important construct in this study, was associated with height, income, education and health. Women who reported ‘a little’ vs. ‘none’ or vs. ‘a great deal’ were taller, had higher incomes and better education, but those with ‘none’ had better health. Importantly, as far as I am aware, religion/religiosity and height have not been previously examined. Replication and validation in other groups are suggested.A possible limitation of this study is that the data are from a cross-section observational study, which may not be sufficient for analyzing changes over time or causal inference. The strengths of this study are the large sample size and reliability and validity of the questionnaire.In conclusion, among relatively healthy U.S. women, 17% Non-White and 83% White, ages 49–79, height and income, and height and education, were not associated. However, taller White women had better jobs, and taller Non-White had better health. In addition, two new results emerged—first, taller Non-White and White women attended clubs/groups more frequently. Second, taller women reported ‘a little’ comfort from religion –they add to the vast literature on height and its relation with human behavior and with well-being. Whether these findings are generalizable globally to diverse populations and a range of demographics– including age, gender, culture, socioeconomics, psychosocial, among others–raise important questions in search of answers.


Physical and operative severity score for the enumeration of mortality and morbidity (POSSUM) scoring system was designed to predict the postoperative morbidity and mortality mainly in general surgery. The purpose of this study was to assess the value of POSSUM scoring system in predicting outcomes of patients undergoing orthopedic surgery, and to do some modifications to make the system more accurate in predicting postoperative complication rates.This is a retrospective clinical study involving 779 patients between April 1, 2009 and September 1, 2010. The postoperative complication rates were predicted by POSSUM, and then compared with the actual morbidity. Logistic regression was taken to improve the POSSUM equation.In the 779 cases, the predicted morbidity was 27.2 % (212 cases) by POSSUM, while the actual morbidity is 8.3 % (65 cases).POSSUM excessively predicted the morbidity of patients undergoing orthopedic surgery, and it could be more accurate with appropriate modification. Of all risk factors, echocardiography ejection fraction showed a close relationship with postoperative complications. Physical and operative severity score for the enumeration of mortality and morbidity (POSSUM) scoring system was first created by Copeland in 1991 . V-POSSUWith the development of medical science, pulmonary function test, echocardiography, ultrasound of lower extremity vascular, ultrasound of carotid, SPECT-CT, etc., were born. Compared to traditional inspection methods, the new medical equipment are more sensitive and accurate. To solve the above problems, we tried to make a new scoring system to predict morbidity and mortality more accurately for orthopedic patients by adding some items in POSSUM scoring system.A total of 779 consecutive patients who had undergone orthopedic surgery between April 1, 2009 and September 1, 2010 were included retrospectively in this study. The study was approved by the ethics committee in our hospital. We included 334 men and 445 women ranging in age from 12 to 96 years with a mean of 61.0 years. Among them, there were 207 cases of the lumbar spine disease, 40 cases of cervical disease, 86 of joint disease, 255 of fracture, 15 of peripheral nerve entrapment, 51 of tumor, 88 of sports injuries, 3 of infectious diseases, and 34 of other diseases. For every included patient, a file was created. All the related data, including patient demographic data , admission/discharge diagnosis, surgeon, surgical approach, physiological scores and operative severity scores of POSSUM, other related indicators , and mortality during period of hospitalization, morbidity occurred within 30 days after surgery, were recorded.POSSUM scoring system consists of 12 physiological scores and 6 operative severity scores, as shown in Table χ2 test. Multivariate analysis was performed using the logistic regression method. P <0.05 was considered statistically significant. The discriminatory power was assessed by the area under the curve (AUC). Values ranging from 0.5 to 0.7 showed a lower diagnostic accuracy, 0.7 to 0.9 represented a moderate diagnostic accuracy, and AUC > 0.9 was a higher diagnostic accuracy.A database involving a total of 779 consecutive patients were established by Epidata 3.1. In order to predict postoperative morbidity or mortality from the lowest risk to the highest risk, 10 risk bands were divided. Each risk band contained the same number of subjects. Expected and observed complications or death were quantified in each band. Then the observed morbidity rate and the expected morbidity rate ratio were calculated (OE = observed complication rate/expected complication rate). SPSS 17.0 software  was used to analyze the data. Missing values were defined as normal (score = 1). Physiologic and operative severity scores were described by mean ± standard deviation. Data was compared by the Within 30 days after surgery, a total of 82 complication cases happened in 65 patients, including 8 cases in myocardial infarction, heart failure, arrhythmia, and other cardiac complications, 19 cases in respiratory failure, pneumonia, and other respiratory complications, 14 cases in cerebral infarction, delirium, and other cerebrovascular complications, 16 cases in wound infection, seroma, infection with implant and bedsores, 8 cases in peptic ulcer, cholecystitis, pancreatitis, and other gastrointestinal complications, 11 cases in lower limb venous thrombosis, 6 cases in urinary tract infections, urinary retention, urinary tract complications. In addition, 2 patients died during the hospital stay.χ2 = 94.88, P < 0.001). In a word, POSSUM overpredicted the morbidity in orthopedic patients. The observed morbidity and the expected morbidity (OE ratio) were shown in Table The average physiologic score was 17.2 ± 4.36, and the average operative severity score was 10.4 ± 3.70, which predicted the probability of complication rate was 27.2 % (212 cases). However, the actual complication rate was 8.3 % (65 cases). Besides, there was a significant difference between the observed complication rate and the expected complication rate, (P <0.001). The ejection fraction was divided into 4 grades: >65 % = 1 score, 55–65 % = 2 score, 40–55 % = 4 score, <40 % = 8 score. Moreover, the 4 grades had a significant correlation with actual morbidity (P <0.001). Therefore, echocardiography ejection fraction had a significant correlation with the prognosis of orthopedic patients.For physiologic score (PSS) and operative severity score (OSS), preoperative comorbidity, serum albumin, serum creatinine, body mass index, time of hospitalization, ejection fraction, pulmonary one second volume, and blood oxygen values with the actual morbidity were related indicators. However, only echocardiographic ejection fraction have a significant correlation with actual morbidity (P for morbidity). The AUC of modified equation was 0.746 and the AUC of original equation was 0.670  = −6.112 + 1.244 × echocardiographic ejection fraction score + 0.096 × PSS + 0.044 × OSS . So, the modified equation had a better predictive ability than the original one, and it was closer to the actual complication rates.In order to test the equation’s accuracy, another 84 patients with hip fracture in our institute were collected. The morbidity was 27.39 % (23 cases) for original equation and 15.07 % (13 cases) for modified equation. However, the actual complication cases were 15 for the 84 hip fracture patients. There was a significant difference between the observed morbidity and expected morbidity  it was a retrospective study, and all the data came from a district teaching hospital’s orthopedic ward. (2) The sample size of the current study was still not large enough to divide into groups by cardiac, respiratory, and other complications, which may have low power. Therefore, multi-disciplinary, multi-center, and large sample size studies are also needed to confirm the new scoring system.In conclusion, POSSUM excessively predicted the morbidity of patients undergoing orthopedic surgery, and it could be more accurate with appropriate modification. However, based on the limits of the current study, large sample, multi-center studies are still needed.


Maximum labeling efficiency was achieved when 1.5 mg ofloxacin was labeled with 10–20 mCi sodium pertechnetate in the presence of 3 mg D-penicillamine, 75 μg SnCl2. In vitro binding and biodistribution in Pseudomonas aeruginosa, Salmonella typhi, and Escherichia coli showed good results. This new complex is efficient for the imaging of infections caused by Gram-positive and Gram-negative bacteria.The employment of radiopharmaceuticals is increasing nowadays for infection imaging and early execution of patients having infectious or inflammatory complaints. The main aim of this study was to discover a novel method for the labeling of ofloxacin with  Many imceutical –8. Amonglability .The radiolabeled antibiotic kits will be considered ideal if it is easily metabolized, has rapid clearance out of body, and has high level of localization at infection site. Certain quinolones that have broad spectrum such as norfloxacin, sitafloxacin, ciprofloxacin, enrofloxacin, ceftriaflaxone, lemofloxacin, difloxacin, pefloxacin, and ceftizoxime are extensively used in the treatment of serious bacterial infection –18.99mTc-ciprofloxacin is an important commercially available radiopharmaceutical that is used as infection imaging agent [ng agent –22. On tng agent –27 and dng agent , 27–29.99mTc such as use of HCl, heating, and instability in serum; thus there is a need to search more efficient, robust method for the development of ofloxacin kits, its radiolabeling, and in vivo studies to sketch the whole profile of the pharmacokinetics and efficacy. Therefore the purpose of the presented work was to develop a simple and well-organized method at room temperature and pressure  to develop 99mTc-ofloxacin, its optimization producing high labeling efficiency and greater stability , and its biodistribution in Salmonella typhi, Pseudomonas aeruginosa, and E. coli infected rabbits.Ofloxacin is very potent member of fluoroquinolone family with broad spectrum against a wide variety of bacterial infections; its structure is shown in μm filter  were used in present study.D-Mannitol , sodium pyrophosphate , octanol , acetone , D-penicillamine , stannous chloride , gentistic acid , ascorbic acid , trichloroacetic acid , saline , ofloxacin , and 0.22 2, amount of antibiotic, pH, and reaction time were studied.Optimization studies were performed to find out the best conditions for the labeling of ofloxacin with technetium. For this purpose SnClμg. It was found that maximum labeling efficiency was obtained at 75 μg; below and above this labeling efficiency was low. All the tests were performed three times (n = 3). Results are summarized in Amount of reducing agent was varied between 25 and 300 Labeling efficiency was analyzed at 0.5–2.5 mg of ofloxacin. It was found that, at 1.5 mg ofloxacin, maximum labeling efficiency was obtained. Results are given in Labeling efficiency (LE) was tested between 2, 3, 4, 5, 6, and 7 pH values and it was concluded that pH 4 was best to get highest value of LE. The results are reported in μg D-penicillamine, and 20 mg D-mannitol were dissolved in 1.0 mL distilled water with continuous stirring (Solution A). Then stannous chloride, 75 μg (15 μL) from a solution having 50 mg SnCl2, and 50 mg Sod/Pot pyrophosphate in 10 mL distilled water were added dropwise to Solution A  from in-house iv)packgen 99mTc-ofloxacin. Paper chromatography (PC) was used to calculate free pertechnetate (99mTcO4−); instant thin layer chromatography (ITLC) was used to calculate the hydrolyzed/reduced technetium (99mTcO2). Counts of 99mTc-ofloxacin were measured through a radioactive detector, that is, Berthold (LB-507) and NaI (Tl) scintillation detector in a Reverse Phase HPLC (RP-HPLC); Perk Elmer (L7200). Nucleosil C-18 column  was used for analytical separations. The solvents variable gradients are 95% A and 5% B at 0 min, 95% A and 5% B at 5 min, 0% A and 100% B at 25 min, and 0% A and 100% B at 30 min with rate of flow 0.5 mL per min. The solvent detail is A = 0.1% tetrafluoroacetic acid in H20 and B = acetonitrile.ITLC, PC, and HPLC were used to assess the radiochemical purity of the Stability studies were performed at room temperature and in fresh human blood serum at 37°C and labeling efficiency was calculated at regular intervals after 1 h, 2 h, 3 h, 4 h, 5 h, and 6 h .μL of both inorganic and organic layers were calculated with well-type (NaI) γ-counter. Partition coefficient values are calculated by mean .Octanol/saline was used as organic and inorganic layers in order to calculate partition coefficient. Counts in 100 P value: Log .Consider the following: LogProtein binding of the labeled complex was measured by incubating it with fresh human blood for 1 h, placing it in a preset water bath (37°C) for ten minutes, centrifuging at 3000 rpm, including equal volume of 10% “trichloroacetic acid” (TCA), and again centrifuged. Afterward both supernatant and residue were counted for radioactivity. Salmonella typhi, Pseudomonas aeruginosa, and E. coli, using cylinder plate method of microbiological assay (used for the potency determination of antibiotics) [It was studied against three bacterial strains, that is,biotics) .99mTc-ofloxacin was injected in the iliac vein of rabbit after valium (1.0 mL) anesthesia. Scintigraphic images were taken at 30 min, 1 h, 2 h, 3 h, and 4 h after injection with Xeleris 2.0523 Gamma camera  of a camera .8 cfu/mL) of Salmonella typhi, Pseudomonas aeruginosa, and E. coli was injected in three different rabbits (thigh muscles). After the initiation of infection and swelling at the infected site of rabbits, 99mTc-ofloxacin (1.0 mL = 2.5 mCi) was injected. Scintigraphy images were taken at 1 h–4 h and 24 h after injection with Infinia Dual head Gamma Camera, that is, Figures For biodistribution study in infected rabbits, 1.0 mL suspension  in the presence of 75 ecreased .99mTc. In PC acetone was used as solvent; small amount of the activity was moved which belonged to free 99mTcO4.HPLC studies of the complex depict that a single compound is formed after this reaction and its retention time was 13.64 min which also matches with the reported value in literature  and yielP = −2.58) and LogP value suggests that dilution of injection will be made with saline.In vitro stability tests at room temperature and in human serum at 37°C showed consistency for 6 h  with no  Escherichia coli, Salmonella typhi, and Pseudomonas aeruginosa showed encouraging results. Biodistribution in normal rabbit showed higher value of activity in kidney and bladder and it is suggested that medicine is excreted mainly through this pathway (T/NT) ratios for Escherichia coli were 2.39 ± 0.17 and 1.8 ± 1, Pseudomonas aeruginosa 3.63 ± 0.16 and 3.13 ± 0.18, and Salmonella typhi 4.47 ± 0.19 and 4.06 ± 0.20  at 4°C and being safe to use with low colloidal amount. Biodistribution studies of the complex in infected rabbits showed hopeful results and it may be used in future in clinical tests for the diagnosis of bone, skin, sinus, lungs, airways, ear, and joint infections caused by the susceptible bacteria.In present investigation,


F. glumosa extract as a diuretic in the treatment of hypertension as claimed by traditional healers. The experiments were performed under the same conditions with two synthetic pharmacological diuretics considered as check (Furosemide and Amiloride hydrochlorothiazide). The aqueous extract leaves of F. glumosa accelerated the elimination of overloaded fluid. At the maximum of diuretic response, urinary osmolarity decreased significantly when compared with controls. The single dose treatment of the aqueous extract leaves of F. glumosa has significantly increased urine volume 24 h after administration of the extract. The stability of aldosterone level, the absence of correlation with the plasma levels of sodium, and the increased clearance of free water in the animals receiving the extract show that increased diuresis and natriuresis moderate elevation are tubular in origin. The increase in Na+, K+, and Cl− induced by the extract caused alkalinization of the urine and showed a strong inhibitory effect of carbonic anhydrase and saluretic. These effects were mainly observed at the dose of 375 mg/kg. These observations confirm the traditional use in the treatment of hypertension and support the importance of the conservation of local knowledge as well as the conservation of Cameroonian biodiversity.Experiments were carried out to validate the use of  Ficus glumosa (Moraceae) is a small- to medium-sized tree, 5–10 m tall. Branches are widely spread, more or less horizontal, often supported by stilt roots. This plant is used in traditional medicine in Cameroon, Senegal, and East Africa for the treatment of edema, hypertension, diabetes, hemorrhoids, rheumatism, and stomatitis [ F. glumosa extracts on gastrointestinal motility were demonstrated in 2012 by Tanko et al. [ Ficus glumosa on fasting blood glucose and serum lipid profile in diabetic rats. Leaves of this tree are also used to cook porridge in Far North of Cameroon and southern Chad. Decoction of the leaves is used as a galactogenic for both women and female animals [ F. glumosa in acute treatment.The overuse in automedication of phytotherapeutic preparations is the main means to cure about 80% population who is unable to get access to manufactured drugs . High blomatitis , 4. The omatitis , 7. One o et al. . Umar et animals . Phytoch animals . The pre F. glumosa were harvested in Ngaoundéré Adamawa region of Cameroon. Then we proceeded to the identification by comparing the harvested plant to specimen no. 60695/HNC deposited at the National Herbarium of Cameroon.The leaves ofWistar rats (250–350 g) of both sexes were used in all experiments. Strains of animals were from Centre Pasteur in Yaoundé. They were reared in the Department of Biological Sciences, Faculty of Sciences (University of Ngaoundéré). The animals were housed under controlled temperature (24 ± 2°C) and relative humidity (45% ± 10%). Moreover, they had free access to food (pellets from LANAVET (Laboratory NVS)) and tap water. The animal handling was under the control of the veterinary surgeon of the Science Veterinary Surgeon and Medical School of the University of Ngaoundéré. Experimental protocols and procedures were approved by the Institutional Animals Care and Use Committee and the research was approved by the Animal Ethics Committee of the University of Ngaoundéré. F. glumosa was steeped in 1 L of distilled water for 12 hours at room temperature. The macerate was filtered through Whatman filter paper No. 3, and the filtrate was concentrated in a rotary evaporator at 40°C for 24 hours. This process repeated several times yielded 11,2 g of concentrated crude extract in the form of an oily paste. The extract was stored at −20°C.One thousand grams of fresh leaves of F. glumosa at increasing doses of 75, 150, 225, 300, and 375 mg/kg was administered to animals at 0.5 mL/100 g body weight. Urine volume was determined 1, 2, 3, 4, 5, 6, and 24 hours after administration of the extract. The animals of control groups were treated with distilled water, Furosemide (2 mg/kg), and Amiloride hydrochlorothiazide (14 mg/kg).In preliminary tests, 1 mL/100 g body weight of distilled water was administered to rats. They were then individually placed in metabolic cages where urine was collected and the volume was measured after 6 hours. The animals excreting at least 40% of the volume of dosing solution were selected for tests. Those having less than 40% excretion were excluded [The animals were sacrificed by decapitation at the end of the experiment. The arteriovenous blood was collected in heparinized tubes and centrifuged. The plasma collected was stored at −20°C for biochemical analysis. The kidney was removed, cleaned of fat material, weighed, and stored at 20°C for biochemical analyzes.2OH = V − Cosm) is positive. Glomerular filtration is determined from the clearance of creatinine. The GFR (glomerular filtration rate) was assessed by creatinine clearance (CreatC). The amount of Na+ and K+ was calculated as a parameter for the saluretic activity. The ratio of Na+/K+ was calculated for the natriuretic activity. To estimate the carbonic anhydrase inhibition activity, the ratio of Cl−/(Na+ + K+) was calculated [Urinary and plasma concentrations of sodium and potassium ions were determined using flame photometry  according to standard methods . Concentlculated .Analytical tests for the identification of different families of metabolites in crude extracts of the leaves were made at IMPM , Cameroon. The procedures described in 1983 by Trease and Evans  were usen = 5). Comparison of means was made using the Student's t-test and one-way ANOVA of Origin Graph software , software version 6.0. P < 0.05, P < 0.01, and P < 0.001 difference was considered significant.The results expressed are the mean ± SEM  Effect of Ficus glumosa Extract on the Urinary Volume. A single dose-response administration of the aqueous extract of F. glumosa  significantly increased (P < 0.05) the volume of urine 24 hours later. However, the urinary excretion was dose dependant (2O) significantly increased to 292.15 ± 4.69 mL/kg/24 h (35.68% increment) at the dose of 225 mg/kg. At the dose of 300 mg/kg, the volume of urine increased by 49.70%. The highest dose (375 mg/kg) induced 82.54% increase of the treated group. The volume of urine went from 231.31 ± 2.38 mL/kg/24 h in control group to 393.04 ± 3.62 mL/kg/24 h in the group treated with the highest dose   . The latthiazide . F. glumosa at the dose of 300 and 375 mg/kg. It was 120 ± 4.11% in animals treated with Furosemide and 88.43 ± 4.41% in those treated with Amiloride hydrochlorothiazide  3 hours after administration significantly (P < 0.05) increased dose-dependently the quantity of urinary excretion. Urine volume increased from 87.44 ± 1.41 mL/kg in controls to 98.14 ± 1.67 mL/kg at the dose of 225 mg/kg, meaning an increase of 12.23% and, respectively, to 133.68 ± 1.22 mL/kg and 136.78 ± 1.08 at doses 300 and 375 mg/kg. The urinary volume of animals treated with Furosemide and Amiloride hydrochlorothiazide meanwhile has increased by 61.87% and 51.04%, respectively  Effect of Aqueous Extract of F. glumosa on Urine Output Index and pH. Diuretic index of the plant extract is lower than that of Furosemide and higher than that of Amiloride hydrochlorothiazide at doses 225 mg/kg, 300 mg/kg, and 375 mg/kg ( F. glumosa showed a significantly (P < 0.05) high urinary pH when compared to the control group ( F. glumosa were higher than the control group (6.1 ± 0.2). Doses of 225 and 300 mg/kg showed significantly increased pH values. However, the pH values (6.6 ± 0.1) of urine of animals treated with the extract were lower than that of rats treated with pharmacological substances (7.0 ± 0.1) (75 mg/kg . At the ol group . The pH 0 ± 0.1) .(1) Effects of F. glumosa on Cumulative Urinary Excretion of Sodium (Na+).The excretion of Na+ excretion increase was 53.92 ± 1.78 mEq/kg/24 h for the dose of 225 mg/kg, 104.24 ± 1.76 mEq/kg/24 h for the dose of 300 mg/kg, and 153.78 ± 2.02 mEq/kg/24 h for the highest dose (375 mg/kg) (P > 0.05) in urinary K+ excretion. Furosemide also induced a significant increase (P < 0.05) from 18.27 ± 1.01 mEq/kg/24 h in the control group to 95.45 ± 4.16 mEq/kg/24 h in the treated group (5 mg/kg) . The Amied group .(2) Effects of F. glumosa on Cumulative Urinary Excretion of Chlorine (Cl−). Aqueous extract of F. glumosa showed a significant increase in the excretion of chloride ions. Excretion increased from 15.27 ± 3.65 mEq/kg/24 h in controls to 50.92 ± 2.35 mEq/kg/24 h at the dose of 225 mg/kg (25 mg/kg . This in25 mg/kg .(3) Effect of F. glumosa on Cumulative Urinary Excretion of Potassium (K+). Urinary potassium excretion was decreased to 20.51  ±  1.29 mEq/kg/24 h in animals treated with Amiloride hydrochlorothiazide (14 mg/kg). Furosemide (2 mg/kg) as well as the extract induced an increase of potassium excretion of 65.99  ±  5.62 mEq/kg/24 h  showed a significant natriuretic activity. A CAI < 0.8 report indicates a strong decrease in diuretic activity [ F. glumosa doses mentioned above shows a CAI > 0.8, indicating a significant diuretic activity. The saluretic dose index 375 is considerable. With a value of 9.12, it is 1.62 times larger than that of Furosemide which is 5.63.activity . ExtractP < 0.05) of 43.54% and 11.25%, respectively, of creatinine and urea in animals that received the extract at dose 375 mg/kg. The animals receiving Furosemide and Amiloride hydrochlorothiazide also show a significant increase (P < 0.05) in the rate of creatinine and urea (25.8% and 6.96% for Furosemide and 22.58% and 15.04% for Amiloride hydrochlorothiazide). Albuminuria increased from 42.8 ± 5.01 g/L in the control group to 44.8  ±  5.06 g/L in animals that received the extract at a dose of 375 mg/kg, meaning an increase of 4.67%. In animals treated with Furosemide and Amiloride hydrochlorothiazide, there was a significant increase (P < 0.05) in the rate of albumin, respectively, of 4.67% and 2.10%. The concentrations of Na+ and K+ ions were significantly increased (P < 0.05), respectively, 509.24% and 182.16% in animals that received the extract at the dose of 375 mg/kg  decreased by 17.91% and 14.48%, at doses of 225 and 300 mg/kg, respectively . A single dose of Furosemide 2 mg/kg was administered orally to rats in search of diuretic action on renal or hepatic insufficiency [+ and Cl− in the ascending branch of Henle loop. It also has a peripheral and independent renal vascular action [The increase of Naecretion , 20. Gloficiency . It actsr action , 23. At r action .P < 0.05) in excretion of K+ compared to Amiloride hydrochlorothiazide (14 mg/kg), suggesting aqueous extract of the leaves of Ficus glumosa does not act as the Amiloride hydrochlorothiazide. It is known that hypervolemia leads to hypertension [ F. glumosa can then be explained by its diuretic properties.Amiloride hydrochlorothiazide 14 mg/kg) induces elimination of unneeded water and salt by the kidneys from the body into the urine . It is a4 mg/kg irtension . The hyp F. glumosa  andwater follows Na+ by osmosis. Optional re-absorption (controlled) is on the level of thedistal circumvented tube (TCD) and collecting tube under the control of the antidiuretichormone or ADH (or vasopressin) secreted by the pituitary gland and indirectly by thealdosterone. The principal substances secreted at the tubular level are K+, H+, ammonia,creatin, penicillin.ubular secretion allows elimination of certain useless substances or ofsurplus in blood and the maintenance of the blood pH by the control of the secretion ofH+. what results by ↑ secretion of ions H+ this involves ↑ pH sanguin and ↓ secretion ofions H+ what results by ↓ pH sanguin. The re-absorption of water proceeds according toan indirect mode , followed by a continuation ofaction of the system renin-angiotensin.Angiotensin II goes stimulates the corticosurenal. The system renin-angiotensin causes ↑ glomerular filtration what results in the Secretionof renin by the cells of the juxta glomerular apparatus. The Formation of angiotensin IIinvolves the vasoconstriction of the efferent small artery, which results by ↑ bloodpressure in the cluster and ↑ filtration. The Secretion of the aldosterone hormone bycorticosurenal gland causes ↑ reabsorption of Na+ on the level of the collecting tubule, ↑reabsorption of water (which follows Na+ by osmosis), ↑ blood volume, and ↑ bloodpressure.Obligatory re-absorption (not controlled) is a consequence of the re-absorption by activetransport of Na