Variations in the age and growth of yellowfin tuna larvae,Thunnus albacares,collected about the Mississippi River plume

Synopsis Eight hundred and one yellowfin tuna larvae ranging from 2.57–7.48 mm SL were collected near the Mississippi River discharge plume in the Gulf of Mexico during July and September, 1987. Larvae were most abundant at intermediate salinities (i.e. frontal waters) where chlorophylla and macrozooplankton displacement values were also highest. Using sagittal otolith microstructure, we estimated larval ages ranging from 3–14 d. These ages were used to back calculate spawning dates from 13–24 July and 22–31 August. Mean absolute individual growth rate (length age^–1) was 0.47 mm d^–1, with the least squares linear regression SL = 1.67 + 0.47 AGE (r² = 0.60, Pr> F = 0.0001) representing the best growth curve. Highest growth occurred at intermediate salinities near 31%, and temperatures near 29° C. There was significant temporal variation in growth, with larvae collected in July growing slower than those from September (0.37 and 0.48 mm d^–1, respectively). The pooled instantaneous daily mortality rate (Z) of the larvae was estimated to be 0.33 d^–1 (0.16 d^–1 in July and 0.41 d^–1 in September). These results show that significant spawning of yellowfin tuna may occur in the northern Gulf of Mexico in the vicinity of the Mississippi River discharge plume, and suggest that larval growth and survival may be enhanced in the plume frontal waters.     

Elevation-Related Variation in Leaf Stomatal Traits as a Function of Plant Functional Type: Evidence from Changbai Mountain,China

Understanding the variation in stomatal characteristics in relation to climatic gradients can reveal the adaptation strategies of plants, and help us to predict their responses to future climate changes. In this study, we investigated stomatal density (SD) and stomatal length (SL) in 150 plant species along an elevation gradient (540–2357 m) in Changbai Mountain, China, and explored the patterns and drivers of stomatal characteristics across species and plant functional types (PFTs: trees, shrubs, and herbs). The average values of SD and SL for all species combined were 156 mm^–2 and 35 µm, respectively. SD was higher in trees (224 mm^–2) than in shrubs (156 mm^–2) or herbs (124 mm^–2), and SL was largest in herbs (37 µm). SD was negatively correlated with SL in all species and PFTs (P<0.01). The relationship between stomatal characteristics and elevation differed among PFTs. In trees, SD decreased and SL increased with elevation; in shrubs and herbs, SD initially increased and then decreased. Elevation-related differences in SL were not significant. PFT explained 7.20–17.6% of the total variation in SD and SL; the contributions of CO₂ partial pressure (), precipitation, and soil water content (SWC) were weak (0.02–2.28%). Our findings suggest that elevation-related patterns of stomatal characteristics in leaves are primarily a function of PFT, and highlight the importance of differences among PFTs in modeling gas exchange in terrestrial ecosystems under global climate change.     

Age and growth of king and Spanish mackerel larvae and juveniles from the Gulf of Mexico and U.S. South Atlantic Bight

Synopsis Sagittal otoliths from 50 king mackerel 2.9–13.0 mm SL and 72 Spanish mackerel 2.8–22.0 mm SL collected off the southeast U.S. were examined whole at 400 × using a compound microscope-video system. Otoliths of both species had visible, presumably daily, growth increments as well as finer subdaily increments. Otolith growth was directly proportional to growth in standard length for king (r² = 0.91) and Spanish mackerel (r² = 0.86). Spanish mackerel were estimated to be 3–15 d old with a mean absolute growth rate (SL/number of growth increments) and 95% confidence interval of 1.15 ± 0.07 mm · d^–1. The least squares linear equation: SL = –1.30 + 1.31 (age in days), with r² = 0.67 and p < 0.001, described the relationship between length and age. There was a significant positive relationship between absolute growth rate and fish length. King mackerel were estimated to be 3–15 d old with a mean absolute growth rate of 0.89 ± 0.06 mm d^–1. The least squares linear equation: SL = 0.37 + 0.82 (age in days), with r² = 0.77 and p < 0.001, best described the relationship between length and age. The relationship between growth rate and fish length was not significant. The growth rate of king mackerel was slightly higher for fish from the Mississippi River plume than from all other locations combined, while Spanish mackerel growth rates were not significantly different.     

Oxygen consumption of engorged nymphs and unfed adults of Hyalomma asiaticum ticks (Acari: Ixodidae) exposed to various photoperiods

The oxygen consumption of engorged nymphs of Hyalomma asiaticum was measured at various intervals after drop-off from mice hosts. Duration of nymphal development to the emergence of adults was 25–32 days at 25°C. The oxygen consumption was high immediately after completing the blood meal (193–248 mm³ g^-1 h^-1 but decreased significantly 18 days later (at 25°C) to 45–65 mm³ g^-1 h^-1. It increased again before ecdysis (81–102 mm³ g^-1 h^-1, and also after ecdysis in freshly moulted adults (177–220 mm³ g^-1 h^-1. The oxygen consumption in 8-month-old adult ticks was very variable ranging from 40–42 to 172 mm³ g^-1 h^-1. Neither engorged nymphs nor unfed adult ticks showed any dependence of their respiratory metabolism on the photoperiodic regimes tested (LD 20:4 and LD 12:12, with or without transfer to an alternative photoperiod after engorgement of nymphs).     

Capillary supply and cross-sectional area of slow and fast twitch muscle fibres in man

Summary The muscles triceps brachii, quadriceps femoris (part vastus lateralis) and soleus were analysed in 6 men and 6 women for fibre composition (% slow twitch, ST-fibres and % fast twitch, FT-fibres), fibre cross sectional areas, and capillarization. Also the fraction of fibres enclosed by their own fibre type was analysed together with the capillary supply of these fibres. Fibre composition was 39(19–60)% ST in m. triceps brachii, 60(29–78)% ST in m. vastus lateralis and 73(49–88)% ST in m. soleus. Fibre areas ranged from 2,320 to 16,667 m² being smallest in m. triceps brachii and largest in m. soleus (p<0.05) and with ST fibres being significantly smaller than FT fibres in some of the muscles. In all muscles the shape of the fibres was elliptical with the larger diameter being about twice the smaller diameter. Capillary density per cross sectional muscle area was not related to the fibre composition and was 379(302–500) cap/mm² in m. triceps brachii, 404(284–529) cap/mm² in m. vastus lateralis and 417(333–592) cap/mm² in m. soleus. However, capillary supply expressed as fibre type area per capillary was up to 40% larger for FT-fibres than for ST-fibres within the same muscle (p<0.05). The capillary supply of enclosed fibres was not different from that of fibres surrounded also by the other fibre type. The results demonstrate that the difference in capillary supply to ST and FT-fibres is less distinct in humans than in other mammals, which is consistent with the metabolic potentials also being more alike.     

Properties of an anion-selective channel from rat colonic enterocyte plasma membranes reconstituted into planar phospholipid bilayers

Summary Vesicles derived from epithelial cells of the colonic mucosa of the rat were fused to planar phospholipid bilayer membranes, revealing spontaneously switching anion-conducting channels of 50 pS conductance (at-30 mV with 200mm Cl^– each side). The equilibrium selectivity series was I^– (1.7)/Br^– (1.3)/Cl^– (1.0)/F^– (0.4)/HCO ₃ ^– (0.4)/Na (<0.11.). Only one dominant open-state conductance could be resolved, which responded linearly to Cl^– concentrations up to 600mm. The singlechannel current-voltage curve was weakly rectifying with symmetrical solutions. When 50 mV were exceeded at the highconductance branch of the curve, switching was arrested in the closed state. At more moderate voltages (±40 mV) kinetics were dominated by one open state of about 35-msec lifetime and two closed states of about 2 and 9-msec lifetime. Of these, the more stable closed state occurred less often. At these voltages one additional closed state of significantly longer lifetime (>0.5 sec) was observed.     

Morphological Alterations in Gastrocnemius and Soleus Muscles in Male and Female Mice in a Fibromyalgia Model

Background

Fibromyalgia (FM) is a chronic musculoskeletal pain disorder, characterized by chronic widespread pain and bodily tenderness and is often accompanied by affective disturbances, however often with unknown etiology. According to recent reports, physical and psychological stress trigger FM. To develop new treatments for FM, experimental animal models for FM are needed to be development and characterized. Using a mouse model for FM including intermittent cold stress (ICS), we hypothesized that ICS leads to morphological alterations in skeletal muscles in mice.

Methods

Male and female ICS mice were kept under alternating temperature (4°C/room temperature [22°C]); mice constantly kept at room temperature served as control. After scarification, gastrocnemius and soleus muscles were removed and snap-frozen in liquid nitrogen–cooled isopentane or fixed for electron microscopy.

Results

In gastrocnemius/soleus muscles of male ICS mice, we found a 21.6% and 33.2% decrease of fiber cross sectional area (FCSA), which in soleus muscle concerns the loss of type IIa and IIx FCSA. This phenomenon was not seen in muscles of female ICS mice. However, this loss in male ICS mice was associated with an increase in gastrocnemius of the density of MIF⁺ (8.6%)-, MuRF⁺ (14.7%)-, Fbxo32⁺ (17.8%)-cells, a 12.1% loss of capillary contacts/muscle fiber as well as a 30.7% increase of damaged mitochondria in comparison with male control mice. Moreover, significant positive correlations exist among densities (n/mm²) of MIF⁺, MuRF⁺, Fbxo32⁺-cells in gastrocnemius/ soleus muscles of male ICS mice; these cell densities inversely correlate with FCSA especially in gastrocnemius muscle of male ICS mice.

Conclusion

The ICS-induced decrease of FCSA mainly concerns gastrocnemius muscle of male mice due to an increase of inflammatory and atrogenic cells. In soleus muscle of male ICS and soleus/gastrocnemius muscles of female ICS mice morphological alterations seem to occur not at all or delayed. The sex-specificity of findings, which is not easily reconciled with the epidemiology of FM (female predominance), implicate that gastrocnemius muscle of male ICS mice should preferentially be used for future investigations with FM. Moreover, we suggest to investigate morphological and/or molecular alterations at different time-points (up to two weeks) after ICS.     

Delay in cART Initiation Results in Persistent Immune Dysregulation and Poor Recovery of T-Cell Phenotype Despite a Decade of Successful HIV Suppression

Background

Successful combination antiretroviral therapy (cART) increases levels of CD4+ T-cells, however this increase may not accurately reflect long-term immune recovery since T-cell dysregulation and loss of T-cell homeostasis often persist. We therefore assessed the impact of a decade of effective cART on immune regulation, T-cell homeostasis, and overall T-cell phenotype.

Methods

We conducted a retrospective study of 288 HIV+ cART-naïve patients initiating therapy. We identified 86 individuals who received cART for at least a decade, of which 44 consistently maintained undetectable plasma HIV-RNA levels throughout therapy. At baseline, participants were classified into three groups according to pre-treatment CD4+ T-cell counts: Group I (CD4<200 cells/mm³); Group II (CD4: 200–350 cells/mm³); Group III (CD4>350 cells/mm³). Outcomes of interest were: (1) CD4+ T-cell count restoration (CD4>532 cells/mm³); (2) normalization of CD4:CD8 T-cell ratio (1.2–3.3); (3) maintenance of CD3+ T-cell homeostasis (CD3: 65%–85% of peripheral lymphocytes); (4) normalization of the complete T-cell phenotype (TCP).

Results

Despite a decade of sustained successful cART, complete T-cell phenotype normalization only occurred in 16% of patients, most of whom had initiated therapy at high CD4+ T-cell counts (>350 cells/mm³). The TCP parameter that was the least restored among patients was the CD4:CD8 T-cell ratio.

Conclusions

Failure to normalize the complete T-cell phenotype was most apparent in patients who initiated cART with a CD4+ T-cell count <200 cells/mm³. The impact of this impaired T-cell phenotype on life-long immune function and potential comorbidities remains to be elucidated.     

Tumor Necrosis Factor-stimulated Gene-6 (TSG-6) Amplifies Hyaluronan Synthesis by Airway Smooth Muscle Cells

We tested the hypothesis that the artificial addition of heavy chains from inter-α-inhibitor to hyaluronan (HA), by adding recombinant TSG-6 (TNF-stimulated gene-6) to the culture medium of murine airway smooth muscle (MASM) cells, would enhance leukocyte binding to HA cables produced in response to poly(I:C). As predicted, the addition of heavy chains to HA cables enhanced leukocyte adhesion to these cables, but it also had several unexpected effects. (i) It produced thicker, more pronounced HA cables. (ii) It increased the accumulation of HA in the cell-associated matrix. (iii) It decreased the amount of HA in the conditioned medium. Importantly, these effects were observed only when TSG-6 was administered in the presence of poly(I:C), and TSG-6 did not exert any effect on its own. Increased HA synthesis occurred during active, poly(I:C)-induced HA synthesis and did not occur when TSG-6 was added after poly(I:C)-induced HA synthesis was complete. MASM cells derived from TSG-6^−/−, HAS1/3^−/−, and CD44^−/− mice amplified HA synthesis in response to poly(I:C) + TSG-6 in a manner similar to WT MASM cells, demonstrating that they are expendable in this process. We conclude that TSG-6 increases the accumulation of HA in the cell-associated matrix, partially by preventing its dissolution from the cell-associated matrix into the conditioned medium, but primarily by inducing HA synthesis.     

Evaluating Total Lymphocyte Count as a Surrogate Marker for CD4 Cell Count in the Management of HIV-Infected Patients in Resource-Limited Settings: A Study from China

Objective

To evaluate the correlation of total lymphocyte count (TLC) and CD4 cell count and the suitability of TLC as a surrogate marker for CD4 cell count of HIV-infected patients in China.

Methods

Usefulness of TLC as a surrogate marker for a CD4 cell count <350 cells/mm³ for HIV-positive patients in China was evaluated by 977 pairs of TLC and CD4 cell count from 977 outpatients. The result was then validated by a literature review which was conducted on 9 relevant articles. Further investigation using the 977 pairs of TLC and CD4 cell count data was done to determine a TLC threshold for predicting a CD4 cell count <500 cells/mm³. Correlation and receiver operating characteristic (ROC) analysis were performed for both CD4 cell counts, and the sensitivity and specificity were computed.

Results

Good correlation was noted between TLC and CD4 count (r = 0.60, 95% CI, 0.56–0.64). TLC obtained a relatively high diagnostic performance (area under ROC curve, 0.80) for predicting a CD4 cell count <350 cells/mm³, with a sensitivity of 0.65 (95% CI, 0.61–0.68) and a specificity of 0.80 (95% CI, 0.75–0.85) at the TLC threshold of 1570 cells/mm³. The literature review suggested that for a CD4 cell count <350 cells/mm³, the optimal TLC threshold was 1500 cells/mm³, which was similar to the figure presented in this observational study. As for predicting a CD4 cell count <500 cells/mm³, TLC obtained a high diagnostic performance (area under ROC curve, 0.82) as well with a sensitivity of 0.70 (95% CI, 0.67–0.73) and a specificity of 0.80 (95% CI, 0.73–0.87).

Conclusions

When considering the antiretroviral therapy for HIV-infected Chinese individuals, total lymphocyte count can be considered as an inexpensive and easily available surrogate marker for predicting two clinically important thresholds of CD4 count of 350 cells/mm³ and 500 cells/mm³.     

Chimeric cells of maternal origin do not appear to be pathogenic in the juvenile idiopathic inflammatory myopathies or muscular dystrophy

IntroductionMicrochimeric cells have been studied for over a decade, with conflicting reports on their presence and role in autoimmune and other inflammatory diseases. To determine whether microchimeric cells were pathogenic or mediating tissue repair in inflammatory myopathies, we phenotyped and quantified microchimeric cells in juvenile idiopathic inflammatory myopathies (JIIM), muscular dystrophy (MD), and noninflammatory control muscle tissues.MethodFluorescence immunophenotyping for infiltrating cells with sequential fluorescence in situ hybridization was performed on muscle biopsies from ten patients with JIIM, nine with MD and ten controls.ResultsMicrochimeric cells were significantly increased in MD muscle (0.079 ± 0.024 microchimeric cells/mm² tissue) compared to controls (0.019 ± 0.007 cells/mm² tissue, p = 0.01), but not elevated in JIIM muscle (0.043 ± 0.015 cells/mm²). Significantly more CD4+ and CD8+ microchimeric cells were in the muscle of patients with MD compared with controls (mean 0.053 ± 0.020/mm² versus 0 ± 0/mm²p = 0.003 and 0.043 ± 0.023/mm² versus 0 ± 0/mm²p = 0.025, respectively). No differences in microchimeric cells between JIIM, MD, and noninflammatory controls were found for CD3+, Class II+, CD25+, CD45RA+, and CD123+ phenotypes, and no microchimeric cells were detected in CD20, CD83, or CD45RO populations. The locations of microchimeric cells were similar in all three conditions, with MD muscle having more microchimeric cells in perimysial regions than controls, and JIIM having fewer microchimeric muscle nuclei than MD. Microchimeric inflammatory cells were found, in most cases, at significantly lower proportions than autologous cells of the same phenotype.ConclusionsMicrochimeric cells are not specific to autoimmune disease, and may not be important in muscle inflammation or tissue repair in JIIM.     

Response of chloride efflux from skeletal muscle ofRana pipiens to changes of temperature and membrane potential and diethylpyrocarbonate treatment

Summary Efflux of³⁶Cl^– from frog sartorius muscles equilibrated in two depolarizing solutions was measured. Cl^– efflux consists of a component present at low pH and a pH-dependent component which increases as external pH increases.For temperatures between 0 and 20°C, the measured activation energy is 7.5 kcal/mol for Cl^– efflux at pH 5 and 12.6 kcal/mol for the pH-dependent Cl^– efflux. The pH-dependent Cl^– efflux can be described by the relationu=1/(1+10n(pK _a -pH)), whereu is the Cl^– efflux increment obtained on stepping from pH 5 to the test pH, normalized with respect to the increment obtained on stepping from pH 5 to 8.5 or 9.0. For muscles equilibrated in solutions containing 150mm KCl plus 120mm NaCl (internal potential about –15 mV), the apparent pK _a is 6.5 at both 0 and 20°C, andn=2.5 for 0°C and 1.5 for 20°C. For muscles equilibrated in solutions containing 7.5mm KCl plus 120mm NaCl (internal potential about –65 mV), the apparent pK _a at 0°C is 6.9 andn is 1.5. The voltage dependence of the apparent pK _a suggests that the critical pH-sensitive moiety producing the pH-dependent Cl^– efflux is sensitive to the membrane electric field, while the insensitivity to temperature suggests that the apparent heat of ionization of this moiety is zero. The fact thatn is greater than 1 suggests that cooperativity between pH-sensitive moieties is involved in determining the Cl^– efflux increment on raising external pH.The histidine-modifying reagent diethylpyrocarbonate (DEPC) applied at pH 6 reduces the pH-dependent Cl^– efflux according to the relation, efflux=exp(–k·[DEPC]·t), wheret is the exposure time (min) to DEPC at a prepared initial concentration of [DEPC] (mm). At 17°C,k ^–1=188mm·min. For temperatures between 10 and 23°C,k has an apparent Q₁₀ of 2.5. The Cl^– efflux inhibitor SCN^– at a concentration of 20mm substantially retards the reduction of the pH-dependent Cl^– efflux by DEPC. The findings that the apparent pK _a is 6.5 in depolarized muscles, that DEPC eliminates the pH-dependent Cl^– efflux, and that this action is retarded by SCN^– supports the notion that protonation of histidine groups associated with Cl^– channels is the controlling reaction for the pH-dependent Cl^– efflux.     

Chloride transport in apical membrane vesicles from bovine tracheal epithelium: Characterization using a fluorescent indicator

Summary Cl transport in apical membrane vesicles derived from bovine tracheal epithelial cells was studied using the Cl-sensitive fluorescent indicator 6-methoxy-N-(3-sulfopropyl) quinolinium. With an inwardly directed 50 mM Cl gradient at 23°C, the initial rate of Cl entry (J _Cl) was increased significantly from 0.32±0.12 nmol · sec^–1 · mg protein^–1 (mean±sem) to 0.50±0.07 nmol · sec^–1 · mg protein^–1 when membrane potential was changed from 0 to +60 mV with K/valinomycin. At 37°C, with membrane potential clamped at 0 mV, there was a 34±7% (n=5) decrease inJ _Cl from a control value of 0.37±0.03 nmol · sec^–1 · mg protein^–1 upon addition of 0.2mm diphenylamine-2-carboxylate. The following did not alterJ _Cl significantly (J _Cl values gives as percent change from control): 50mm cis Na (–1±5%), 0.1mm furosemide (–3±4%), 0.1mm furosemide in the presence of 50mm cis Na (–5±2%), 0.1mm H₂DIDS (–18±9%), a 1.5 pH unit inwardly directed H gradient (–7±7%), and 0.1mm H₂DIDS in the presence of a 1.5 unit pH gradient (4±18%). With inward 50mm anion gradients, the initial rates of Br and I entry (J _Br andJ ₁, respectively) were not significantly different fromJ _Cl.J _Cl was a saturable function of Cl concentration with apparentK _d of 24mm and apparentV _max of 0.54 nmol · sec^–1 · mg protein^–1. Measurement of the temperature dependence ofJ _Cl yielded an activation energy of 5.0 kcal/mol (16–37°C). These results demonstrate that Cl transport in tracheal apical membrane vesicles is voltage-dependent and inhibited by diphenylamine-2-carboxylate. There is no significant contribution from the Na/K/2Cl, Na/Cl, or Cl/OH(H) transporters. The conductive pathway does not discriminate between Cl, Br, and I and is saturable. The low activation energy supports a pore-type mechanism for the conductance.     

Availability of fluoride to plants grown in contaminated soils

Two pot experiments were carried out to study uptake of fluoride (F) in clover and grasses from soil. Fluoride concentrations in t Trifolium repens (white clover) and t Lolium multiflorium (ryegrass) were highly correlated with the amounts of H₂O– and 0.01 t M CaCl₂–extractable F in soil when increasing amounts of NaF were added to two uncontaminated soils (r=0.95–0.98, t p<0.001). The amounts of H₂O– or 0.01 t M CaCl₂–extractable F did not explain the F concentrations to a similar extent in t Agrostis capillaris (common bent) grown in 12 soils (Cambic Arenosols) collected from areas around the Al smelters at Å: rdal and Sunndal in Western Norway (r=0.68–0.78). This may be due to variation in soil pH and other soil properties in the 12 soils. Soil extraction with 1 t M HCl did not estimate plant–available F in the soil as well as extraction with H₂O or 0.01 t M CaCl₂. Fluoride and Al concentrations in the plant material were positively correlated in most cases. Fluoride and Ca concentrations in the plant material were negatively correlated in the first experiment. No consistent effects were found on the K or Mg concentrations in the plant material. The F accumulation in clover was higher than in the grasses. The uptake from soil by grasses was relatively low compared to the possible uptake from air around the Al smelters. The uptake of F in common bent did not exceed the recommended limit for F contents in pasture grass (30 mg kg^–1) from soil with 0.5–28 mg F(H₂O) kg^–1 soil. The concentration in ryegrass was about 50 mg F kg^–1 when grown in a highly polluted soil (28 mg F(H₂O) kg^–1 soil). Concentrations in clover exceeded 30 mg F kg^–1 even in moderately polluted soil (1.3–7 mg F(H₂O) kg^–1 soil). Liming resulted in slightly lower F concentrations in the plant material.     

T Lymphocyte Density and Distribution in Human Colorectal Mucosa,and Inefficiency of Current Cell Isolation Protocols

Mucosal tissues are critical immune effector sites containing complex populations of leukocytes in a tissue microenvironment that remains incompletely understood. We identify and quantify in human distal colorectal tissue absolute mucosal CD3⁺ lymphocytes, including CD4⁺ and CD8⁺ subsets, by direct visualization using immunohistochemistry (IHC), immunofluorescence (IF), and an automated counting protocol (r²=0.90). Sigmoid and rectal mucosal tissues are both densely packed with T lymphocytes in the mucosal compartment. Both compartments had similar densities of CD3⁺ T lymphocytes with 37,400 ± 2,801 cells/mm³ and 33,700 ± 4,324 cell/mm³, respectively. Sigmoid mucosa contained 57% CD3⁺CD4⁺ and 40% CD3⁺CD8⁺ T lymphocytes which calculates to 21,300 ± 1,476/mm³ and 15,000 ± 275/mm³ T lymphocytes, respectively. Rectal mucosa had 57% CD3⁺CD4⁺ and 42% CD3⁺CD8⁺ or 21,577 ± 332, and 17,090 ± 1,206 cells/mm³, respectively. By comparison, sigmoid mucosal biopsies subjected to conventional collagenase digestion, mononuclear cell (MMC) isolation and staining for flow cytometry yielded 4,549 ± 381/mm³ and 2,708 ± 245/mm³ CD4+ and CD8+ T lymphocytes. These data suggest only ~20.7% recovery compared to IHC results for these markers. Further studies will determine if this reflects a selective bias in only CD3⁺, CD4⁺ and CD8⁺ T cells or can be generalized to all flow-analyzed cells from mucosal tissues for phenotyping and functional testing.     

When to Start Antiretroviral Therapy in Children Aged 2–5 Years: A Collaborative Causal Modelling Analysis of Cohort Studies from Southern Africa

Background

There is limited evidence on the optimal timing of antiretroviral therapy (ART) initiation in children 2–5 y of age. We conducted a causal modelling analysis using the International Epidemiologic Databases to Evaluate AIDS–Southern Africa (IeDEA-SA) collaborative dataset to determine the difference in mortality when starting ART in children aged 2–5 y immediately (irrespective of CD4 criteria), as recommended in the World Health Organization (WHO) 2013 guidelines, compared to deferring to lower CD4 thresholds, for example, the WHO 2010 recommended threshold of CD4 count <750 cells/mm³ or CD4 percentage (CD4%) <25%.

Methods and Findings

ART-naïve children enrolling in HIV care at IeDEA-SA sites who were between 24 and 59 mo of age at first visit and with ≥1 visit prior to ART initiation and ≥1 follow-up visit were included. We estimated mortality for ART initiation at different CD4 thresholds for up to 3 y using g-computation, adjusting for measured time-dependent confounding of CD4 percent, CD4 count, and weight-for-age z-score. Confidence intervals were constructed using bootstrapping.The median (first; third quartile) age at first visit of 2,934 children (51% male) included in the analysis was 3.3 y (2.6; 4.1), with a median (first; third quartile) CD4 count of 592 cells/mm³ (356; 895) and median (first; third quartile) CD4% of 16% (10%; 23%). The estimated cumulative mortality after 3 y for ART initiation at different CD4 thresholds ranged from 3.4% (95% CI: 2.1–6.5) (no ART) to 2.1% (95% CI: 1.3%–3.5%) (ART irrespective of CD4 value). Estimated mortality was overall higher when initiating ART at lower CD4 values or not at all. There was no mortality difference between starting ART immediately, irrespective of CD4 value, and ART initiation at the WHO 2010 recommended threshold of CD4 count <750 cells/mm³ or CD4% <25%, with mortality estimates of 2.1% (95% CI: 1.3%–3.5%) and 2.2% (95% CI: 1.4%–3.5%) after 3 y, respectively. The analysis was limited by loss to follow-up and the unavailability of WHO staging data.

Conclusions

The results indicate no mortality difference for up to 3 y between ART initiation irrespective of CD4 value and ART initiation at a threshold of CD4 count <750 cells/mm³ or CD4% <25%, but there are overall higher point estimates for mortality when ART is initiated at lower CD4 values. Please see later in the article for the Editors'' Summary     

The relative importance of different ciliate taxa in the pelagic food web of lake constance

Abundance, biovolume, and species composition of pelagic ciliates in Lake Constance were recorded over two annual cycles (1987/88). Production was estimated from mean annual biovolumes and size-specific growth rates obtained from the literature. Cell concentrations and biovolumes ranged from 0.1 to 120 cells ml^–1 and from 3 to 1,200 mm³ m^–3, respectively. Mean annual values were, respectively, 6.8 cells ml^–1 and 94 mm³ m^–3 in 1987, and 12.0 cells ml^–1 and 130 mm³ m^–3 in 1988. In both years, prostome nanociliates (<20m) dominated numerically, while strobiliids in the size range 20–35m contributed most significantly to ciliate production. Ciliate community production, according to a crude calculation, yielded approximately 10–15 g C m^–2 year^–1.     

CD4 and Viral Load Dynamics in Antiretroviral-Na?ve HIV-Infected Adults from Soweto,South Africa: A Prospective Cohort

BackgroundCD4 count is a proxy for the extent of immune deficiency and declines in CD4 count are a measure of disease progression. Decline in CD4 count is an important component: for estimating benefits of ARV treatment; for individual level counselling on the rapidity of untreated disease progression and prognosis; and can be used in planning demand for health services. Our objective is to report CD4 decline and changes in viral load (VL) in a group of HIV-infected adults enrolled in a randomized trial of preventive treatment for TB in South Africa where clade C infection predominates.MethodsHIV-infected, tuberculin skin test positive adults who were not eligible for antiretroviral (ARV) treatment were randomized to a trial of preventive treatment from 2003–2005. VL and CD4 count were assessed at enrollment and CD4 counts repeated at least annually. During follow-up, individuals whose CD4 counts decreased to <200 cells/mm³ were referred for antiretroviral therapy (ART) and were analytically censored.Results1106 ARV naïve adults were enrolled. Their median age was 30 years and male to female ratio was 1∶5. Median baseline CD4 count was 490 cells/mm³ (IQR 351–675). The overall mean decline in CD4 count was 61 cells/mm³ per annum. Adjusting for age, gender, baseline hemoglobin, smoking and alcohol use had little impact on the estimate of CD4 decline. However, VL at baseline had a major impact on CD4 decline. The percent decline in CD4 count was 13.3% (95% CI 12.0%, 14.7%), 10.6% (95% CI 8.8%, 12.4%), and 13.8% (95% CI 12.1%, 15.5%) per annum for baseline VLs of <10,000 (N = 314), 10,001–100,000 (N = 338), >100,000 (N = 122) copies/ml.ConclusionsOur data suggests that six and a half years will elapse for an individual''s CD4 count to decline from 750 to 350 cells/mm³ in the absence of ART.     

The Effect of Bathing Solution Tonicity on Resting Tension in Frog Muscle Fibers

Resting tension and short-range elastic properties of isolated twitch muscle fibers of the frog have been studied while bathed by solutions of different tonicities. Resting tension in isotonic solution at 2.3-µm sarcomere spacing averaged 0.46 mN·mm^-2 and was proportional to the fiber cross-section area. Hypertonic solutions, containing 0.1–0.5 mM tetracaine to block contracture tension, caused a small sustained tension increase, which was proportional to the fiber cross-section area and which reached 0.9 mN·mm^-2 at two times normal tonicity (2T). Further increases in tonicity caused little increase in tension. Hypotonic solutions decreased tension. Thus, tension at 2.3 µm is a continuous, direct function of tonicity. The dependence of tension on tonicity lessened at greater sarcomere lengths. At 3.2 µm either a very small rise or, in some fibers, a fall in tension resulted from an increase in tonicity. Hypertonic solutions also decreased the tension of extended sarcolemma preparations. In constant-speed stretch experiments the elastic modulus, calculated from the initial part of the stretch response, rose steeply with tonicity over the whole range investigated (1–2.5T). The results show that tension and stiffness of the short-range elastic component do not increase in parallel in hypertonic solutions.     

The Virological and Immunological Characteristics of the HIV-1-Infected Population in Brazil: From Initial Diagnosis to Impact of Antiretroviral Use

BackgroundImmunological and virological status of HIV-infected individuals entering the Brazilian public system over time was analyzed. We evaluated the impact of ART on virological, immunological and antiretroviral resistance over time.MethodsCD4+ T cell counts, viral loads and genotypes from patients over 13 years old from 2001–2011 were analyzed according to demographic data. We compared groups using parametric t-tests and linear regression analysis in the R statistical software language.ResultsMean baseline CD4+ T cell counts varied from 348 (2003) to 389 (2009) and was higher among women (p = 1.1 x 10⁻⁸), lower in older patients (p< 1 x 10⁻⁸) and lower in less developed regions (p = 1.864 x 10⁻⁵). Percentage of treated patients with undetectable viral loads increased linearly from 46% (2001) to 77% (2011), was lower among women (p = 2.851 x 10⁻⁶), younger ages (p = 1 x 10⁻³), and in less developed regions (p = 1.782 x 10⁻⁴). NRTI acquired resistance was 86% in 2001–3 and decreased over time. NNRTI resistance increased from 2001-3(50%) to 2006–9 (60%), PI resistance decreased from 2001–3 (60%) to 2009 (40%), and 3-class resistance was stable over time around 25%. Subtype prevalence comprised B (75.3%), B/F recombinants (12.2%), C (5.7%), F (5.3%) and B/C recombinants (1.5%), with regional variations. Three-class resistance was 26.5% among Bs, 22.4% among Fs and 17.2% among Cs.ConclusionsHIV diagnosis occurs late, especially among elderly Brazilians. Younger individuals need special attention due to poor virological response to treatment. Antiretroviral Resistance profile is subtype related.