Effect of a CCR5 inhibitor on viral loads in macaques dual-infected with R5 and X4 primate immunodeficiency viruses

Human immunodeficiency virus type 1 (HIV-1) fusion with its target cells is initiated by sequential interactions between its envelope glycoprotein, CD4, and a co-receptor, usually CCR5 or CXCR4. Small molecules that bind to CCR5 and prevent its use by R5 HIV-1 strains are now being developed clinically as antiviral drugs. To test whether a block to CCR5 promotes the replication of viruses that enter cells via CXCR4 and are associated with accelerated disease progression, we administered a small molecule CCR5 inhibitor, CMPD 167, to three macaques dual-infected with both R5 (SIVmac251) and X4 (SHIV-89.6P) viruses. CMPD 167 caused a rapid and substantial (on average, 50-fold) suppression of R5 virus replication in each animal. In two of the animals, but not in the third, a rapid, transient, 8- to 15-fold increase in the amount of plasma X4 virus occurred. In neither animal was the increase in X4 viral load sustained throughout therapy, however. These observations may have relevance for the development of CCR5 inhibitors for treatment of HIV-1 infection of humans.     

Inclusion of a non-immunoglobulin binding protein in two-site ELISA for quantification of human serum proteins without interference by heterophilic serum antibodies

Measurement of human serum molecules with two-site ELISA can be biased by the presence of human heterophilic anti-animal immunoglobulin antibodies (HAIA) that cause false-positive signals by cross-linking the monoclonal (mAb) and/or polyclonal antibodies (pAb) used for the pre- (capture) and post-analyte steps (detection). To evaluate a novel ELISA format designed to avoid interference by HAIA, a target-specific non-immunoglobulin (Ig) affinity protein (affibody) was used to replace one of the antibodies. First, a human IgA-binding affibody (Z(IgA)) selected by phage display technology from a combinatorial library of a single Staphylococcus aureus protein A domain was used. The detection range of IgA standard using an ELISA based on Z(IgA) for capture and goat pAb against IgA (pAb(IgA)) for detection was comparable with that of using pAb(IgA) for both capture and detection. Secondly, another affibody (Z(Apo)) was combined with mAb and used to detect recombinant human apolipoprotein A-1. The affibody/antibody ELISAs were also used to quantify human serum levels of IgA and apolipoprotein A1. To verify that human serum did not cause false-positive signals in the affibody/antibody ELISA format, the ability of human serum to cross-link affibodies, mAb (mouse or rat) and/or pAb (goat) displaying non-matched specificities was assessed; affibodies and antibodies were not cross-linked whereas all combinations of mAb and/or pAb were cross-linked. The combination of affibodies and antibodies for analysis of human serum molecules represents a novel two-site ELISA format which precludes false-positive signals caused by HAIA.     

Recovery from short term intense exercise: Its relation to capillary supply and blood lactate concentration

Summary Muscle force recovery from short term intense exercise was examined in 16 physically active men. They performed 50 consecutive maximal voluntary knee extensions. Following a 40-s rest period five additional maximal contractions were executed. The decrease in torque during the 50 contractions and the peak torque during the five contractions relative to initial torque were used as indices for fatigue and recovery, respectively. Venous blood samples were collected repeatedly up to 8 min post exercise for subsequent lactate analyses. Muscle biopsies were obtained from m. vastus lateralis and analysed for fiber type composition, fiber area, and capillary density. Peak torque decreased 67 (range 47–82%) as a result of the repeated contractions. Following recovery, peak torque averaged 70 (47–86%) of the initial value. Lactate concentration after the 50 contractions was 2.9±1.3 mmol·l⁻¹ and the peak post exercise value averaged 8.7±2.1 mmol·l⁻¹. Fatigue and recovery respectively were correlated with capillary density (r=−0.71 and 0.69) but not with fiber type distribution. A relationship was demonstrated between capillary density and post exercise/peak post exercise blood lactate concentration (r=0.64). Based on the present findings it is suggested that lactate elimination from the exercising muscle is partly dependent upon the capillary supply and subsequently influences the rate of muscle force recovery. Dr. Tesch was on leave from Department of Clinical Physiology, Karolinska Hospital, Stockholm, Sweden     

Contraction-specific differences in maximal muscle power during stretch-shortening cycle movements in elderly males and females

Elderly people (age 75 years; n=48 males and 34 females) were studied in order to elucidate gender differences in elderly subjects on the determinants of muscle power (force and velocity) during a stretch-shortening cycle. All subjects performed three maximal counter-movement vertical jumps using both legs, on a force platform (Kistler 9281 B). The eccentric (Ep) and concentric (Cp) phases of the jumps were analyzed. The Ep was further divided into an acceleration phase (Ep_acc: from the start of the downward movement to the maximal negative velocity) and deceleration phase (Ep_dec: from the maximal negative velocity to the end of the downward movement). Jump height for the men was higher than for the women (P < 0.001). During both Ep_acc and Ep_dec no significant differences were observed between males and females in force and power generation. However, the men had a higher peak muscle power during the Cp, which may be explained exclusively by the velocity determinant (P < 0.001). No specific gender-related strategy appeared to influence the motor pattern of the movement. The comparable eccentric force generation of the leg extensors in both genders suggests a similar ability to cope with eccentric muscle actions during everyday activities. In contrast, the marked lower capacity for concentric contractions in women may result in an impaired performance, especially in activities where intense and rapid movements are essential, for example when reversing a forward fall. This may be one reason why elderly women are more prone to falls than are elderly men. Accepted: 19 September 2000     

Specifically associated PCR products probed by coincident detection of two-color cross-correlated fluorescence intensities in human gene polymorphisms of methylene tetrahydrofolate reductase at site C677T: a novel measurement approach without follow-up mathematical analysis

Whole blood samples of known methylene tetrahydrofolate reductase (MTHFR) genotypes from 24 individuals were examined at site C677T. Their amplified DNA products were assessed by two-color fluorescence cross-correlation measurements and agarose gel electrophoresis/capillary gel electrophoresis. DNA subpopulations were identified which were not associated with the proper genotype by primer combinations and cycling conditions called multiplexes. We confirmed that DNA analysis by two-color fluorescence cross-correlation measurements allowed the detection of fluorescence signals specifically associated with the proper genotypes in a mixture of amplified nontarget DNA molecules without DNA sizing. The measurement approach does not require complex, follow-up mathematical analysis and is applicable to any single nucleotide polymorphisms. The simple immunogenetic model showed how the approach works to reveal specific DNA target by preventing detection of nontarget DNA. Under those experimental conditions, a new ultrasensitive, and specific method for clinical immunologists is born.     

Two-dimensional acoustic attenuation mapping of high-temperature interstitial ultrasound lesions

Acoustic attenuation change in biological tissues with temperature and time is a critical parameter for interstitial ultrasound thermal therapy treatment planning and applicator design. Earlier studies have not fully explored the effects on attenuation of temperatures (75-95 degrees C) and times (5-15 min) common in interstitial ultrasound treatments. A scanning transmission ultrasound attenuation measurement system was devised and used to measure attenuation changes due to these types of thermal exposures. To validate the approach and to loosely define expected values, attenuation changes in degassed ex vivo bovine liver, bovine brain and chicken muscle were measured after 10 min exposures in a water bath to temperatures up to 90 degrees C. Maximum attenuation increases of approximately seven, four and two times the values at 37 degrees C were measured for the three tissue models at 5 MHz. By using the system to scan over lesions produced using interstitial ultrasound applicators, 2D contour maps of attenuation were produced. Attenuation profiles measured through the centrelines of lesions showed that attenuation was highest close to the applicator and decreased with radial distance, as expected with decreasing thermal exposure. Attenuation values measured in profiles through lesions were also shown to decrease with reduced power to the applicator. Attenuation increases in 2D maps of interstitial ultrasound lesions in ex vivo chicken breast, bovine liver and bovine brain were correlated with visible tissue coagulation. While regions of visible coagulation corresponded well to contours of attenuation increase in liver and chicken, no lesion was visible under the same experimental conditions in brain, due primarily to the heterogeneity of the tissue. Acoustic and biothermal simulations were employed to show that attenuation models taking into account these attenuation changes at higher temperatures and longer times were better able to fit experimental data than previous models. These simulations also indicated that the characterization of tissue acoustic and thermal properties over a large range of temperatures is critical for accurate treatment planning or design studies involving high-temperature interstitial ultrasound.     

Follow-up on mental illness in medical inpatients: health care use and self-rated health and physical fitness

Consecutively admitted internal medical inpatients (N=294) who were psychiatrically assessed with the Schedules for Clinical Assessment in Neuropsychiatry in a two-phase design were followed up in a review of public files on their use of medical care over 18 months. Self-rated outcome was assessed from health and fitness ratings at admission and after 1 year. ICD-10 mental disorders had a statistically significant impact on the risk (odds ratio) of high use (above the 80th percentile) of primary care, as did ICD-10 anxiety/depression, and worry about illness (as assessed by the Whiteley-7 Scale). The authors found a less-than-significant tendency for mental illness to influence the use of inpatient admissions and self-rated outcome.     

The effect of a high calory diet on hormonal changes in young men during prolonged physical strain and sleep deprivation

Summary Major changes occur in the serum level of several hormones during 5 days of heavy and continuous physical activities, with less than a total of 2 h of sleep. The present investigation was designed to evaluate the importance of caloric deficiency, energy requirement being about 8,000–10,000 kcal/24 h. A comparison between well fed subjects and those with food deprivation revealed significantly higher levels of (T₃) triiodothyronin, insulin and thyroid stimulating hormone (TSH) in the well-fed subjects, who also had lower levels of growth hormone (hGH) and cortisol, whereas no difference was found between the two groups for thyroxin (T₄). Increased levels were found for T₃ and T₄ in both groups during the first day of activity, with a concomitant decrease in TSH and a subsequent decrease of T₄ during the next 2 days. T₃ decreased only in the low-calory group whereas increased levels were found in the iso-calory group throughout the course. The resting levels of insulin decreased during the course in the low-calory group whereas it increased in the iso-calory group. High levels were maintained throughout the course for hGH. Cortisol showed high levels just before the start of the course and then decreased from day 2 to day 4. No difference was found between the morning and evening levels for cortisol, indicating disappearance of the circadian rhythm. The present investigation has shown that energy deficiency during prolonged physical strain is responsible for the decreased serum levels of T₃ and insulin and may contribute to the decrease in TSH and the increase in hGH and cortisol.     

Naloxone-provoked vaso-vagal response to head-up tilt in men

A double-blind paired protocol was used to evaluate, in eight male volunteers, the effects of the endogenous opiate antagonist naloxone (NAL; 0.05 mg· kg^–1) on cardiovascular responses to 50° head-up tilt-induced central hypovolaemia. Progressive central hypovolaemia was characterized by a phase of normotensive-tachycardia followed by an episode of hypotensive-bradycardia. The NAL shortened the former from 20 (8–40) to 5 (3–10) min (median and range; (P < 0.02). Control head-up tilt increased the means of thoracic electrical impedance [from 35.8 (SEM 2.1) to 40.0 (SEM 1.8) ; P < 0.01 of heart rate [HR; from 67 (SEM 5) to 96 (SEM 8) beats · min^–1, P < 0.02], of total peripheral resistance [TPR; from 25.5 (SEM 3.2) to 50.4 (SEM 10.5)mmHg min 1^–1,P < 0.05] and of mean arterial pressure [MAP; from 96 (SEM 2) to 101 (SEM 2)mmHg, P < 0.02]. Decreases were observed in stroke volume [from 65 (SEM 12) to 38 (SEM 9) ml, P < 0.01], in cardiac output [from 3.7 (SEM 0.7) to 2.5 (SEM 0.5) 1 · mint, P < 0.01], in pulse pressure [from 55 (SEM 4) to 37 (SEM 3)mmHg, P < 0.01] and in central venous oxygen saturation [from 73 (SEM 2) to 59 (SEM 4)%, P < 0.01]. During NAL, mean HR increased from 70 (SEM 3); n.s. compared to control) to only 86 (SEM 9) beats · min^–1 (P < 0.02 compared to control) and MAP remained stable. The episode of hypotensive-bradycardia appeared as mean control HR decreased to 77 (SEM 7)beats · min^–1, TPR to 31.4(SEM 7.7)mmHg · min · 1^–1 and MAP to 60 (SEM 5)mmHg (P < 0.01), and the volunteers were tilted supine. Cardiovascular effects of naloxone on central hypovolaemia included a reduced elevation of HR and blood pressures and provocation of the episode of hypotensive-bradycardia.