Excessive innate immune system activation and inflammation during pregnancy can lead to organ injury and dysfunction and preeclampsia (PE); however, the molecular mechanisms involved are unknown. We tested the hypothesis that Toll-like receptor (TLR) activation induces major histocompatibility complex (MHC) class II invariant chain peptide (CLIP) expression on immune cells, makes them pro-inflammatory, and are necessary to cause PE-like features in mice. Treatment with VG1177, a competitive antagonist peptide for CLIP in the groove of MHC class II, was able to both prevent and treat PE-like features in mice. We then determined that γ–δ T cells are critical for the development of PE-like features in mice since γ–δ T-cell knockout mice, like CLIP deficient mice, are resistant to developing PE-like features. Placentas from women with PE exhibit significantly increased levels of γ–δ T cells. These preclinical data demonstrate that CLIP expression and activated γ–δ T cells are responsible for the development of immunologic PE-like features and that temporarily antagonizing CLIP and/or γ–δ T cells may be a therapeutic strategy for PE.

Pre-eclampsia, the development of hypertension and proteinuria or end-organ damage during pregnancy, is a leading cause of both maternal and fetal morbidity and mortality, and there are no effective clinical treatments for pre-eclampsia aside from delivery. The development of pre-eclampsia is characterized by maladaptation of the maternal immune system, excessive inflammation and endothelial dysfunction. We have reported that detection of extracellular RNA by the Toll-like receptors (TLRs) 3 and 7 is a key initiating signal that contributes to the development of pre-eclampsia. PLacental eXpanded (PLX-PAD) cells are human placenta-derived, mesenchymal-like, adherent stromal cells that have anti-inflammatory, proangiogenic, cytoprotective and regenerative properties, secondary to paracrine secretion of various molecules in response to environmental stimulation. We hypothesized that PLX-PAD cells would reduce the associated inflammation and tissue damage and lower blood pressure in mice with pre-eclampsia induced by TLR3 or TLR7 activation. Injection of PLX-PAD cells on gestational day 14 significantly decreased systolic blood pressure by day 17 in TLR3-induced and TLR7-induced hypertensive mice (TLR3 144–111 mmHg; TLR7 145–106 mmHg; both P <0.05), and also normalized their elevated urinary protein:creatinine ratios (TLR3 5.68–3.72; TLR7 5.57–3.84; both P <0.05). On gestational day 17, aortic endothelium-dependent relaxation responses improved significantly in TLR3-induced and TLR7-induced hypertensive mice that received PLX-PAD cells on gestational day 14 (TLR3 35–65%; TLR7 37–63%; both P <0.05). In addition, markers of systemic inflammation and placental injury, increased markedly in both groups of TLR-induced hypertensive mice, were reduced by PLX-PAD cells. Importantly, PLX-PAD cell therapy had no effects on these measures in pregnant control mice or on the fetuses. These data demonstrate that PLX-PAD cell therapy can safely reverse pre-eclampsia-like features during pregnancy and have a potential therapeutic role in pre-eclampsia treatment.

1. The functional properties of tibialis anterior muscles of normal adult (C57BL/10) and age-matched dystrophin-deficient (C57BL/10 mdx ) mice have been investigated in situ. Comparisons were made between tibialis anterior muscle strength, rates of force development and relaxation, force-frequency responses and fatiguability. Subjecting mdx and C57 muscles to a regimen of eccentric exercise allowed the hypothesis to be tested that dystrophin-deficient muscles are more susceptible to exercise-induced muscle damage. 2. mdx muscles were, on average, 30% stronger than C57 muscles and almost 80% heavier, but both had similar muscle lengths. Thus, although mdx muscles were stronger in absolute terms, their estimated force per unit cross-sectional area was significantly less than that of C57 muscles. 3. The force-frequency relationships of C57 and mdx muscles differed in that whilst, at 40 Hz, the former developed 70% of the force developed at 100 Hz, the latter developed only 55% of the maximal force. Twitch force was normal in mdx muscles, but contraction time was shortened, and the consequent reduction in fusion frequency probably explains the force-frequency differences observed between the two groups. 4. mdx muscles were less fatiguable than normal muscles when stimulated repeatedly at a frequency of 40 Hz. It is possible that the lower relative force at 40 Hz in mdx muscles entailed a lower energy demand and thus a slower rate of fatigue than seen in normal muscles. 5. Eccentrically exercised C57 muscles showed a large loss of maximal force for up to 12 days after exercise. Maximal force loss occurred 3 days after exercise (55% of non-exercised tibialis anterior muscle), which also corresponded with the period of greatest fibre necrosis. C57 muscles showed a significantly reduced 40 Hz/100 Hz force-frequency ratio at 1 and 3 days after exercise. This was primarily due to a reduced twitch amplitude rather than to a change in the time course of the twitch. It is unlikely, therefore, that the altered contractile characteristics of mdx muscle were a result of the presence of damaged but otherwise normal fibres. 6. C57 and mdx tibialis anterior muscles displayed similar degrees of force loss after exercise. Furthermore, the rate of recovery after the nadir of force loss was very similar for the two groups. By 12 days after exercise, force recovered to 76% and 80% of control in C57 and mdx muscles, respectively. Our findings do not support the hypothesis that dystrophin-deficient muscle is more susceptible to exercise-induced muscle damage.

1. The effect of muscle length on the development of muscle pain and fatigue has been studied. 2. Eight normal young adults performed maximal eccentric contractions of the elbow flexors. The muscles of one arm were exercised at short length, and the contralateral muscle at long length. Each contraction lasted approximately 1 s, and was repeated once every 10 s for 30 min. 3. Muscle strength and frequency-force characteristics were measured from isometric contractions before, immediately after and at 24 h intervals for the next 4 days. Muscle tenderness was assessed daily. 4. The muscle strength was reduced by approximately 10% by exercise at short length, and by 30% by exercise at long length. 5. The 20:100 ratio (force generated by stimulation at 20 Hz/force generated at 100 Hz) fell by 30% after exercise at short length and had recovered after 24 h. Exercise at long length reduced this ratio by 65% and the muscles had not fully recovered 4 days later. 6. Muscle pain developed after both exercise regimens, but was slightly worse after that at long length. 7. It is concluded that there is a length-dependent component in the development of pain and fatigue after eccentric exercise, which had previously been thought to be caused solely by high force generation.

1. The involvement of extracellular calcium in experimental muscle damage has been studied in an isolated mouse soleus muscle preparation. 2. The enzyme efflux and ultrastructural damage seen after excessive contractile activity were markedly reduced when the extracellular calcium was withdrawn. Low extracellular calcium also protected against the large enzyme efflux seen after treatment with low concentrations of detergent. 3. Treatment of the muscle with the calcium ionophore A 23187 caused significant release of enzyme from the muscle. 4. Nifedipine did not prevent the enzyme release after stimulation and although in some circumstances verapamil appeared to have some protective effect this was probably due to a local anaesthetic action on the muscle and not to any specific effect on calcium movement. 5. It is concluded that extracellular calcium is important in mediating at least the two forms of muscle damage studied here.

1. Activities for the oxidation of palmitoyl-carnitine, of palmitoyl-CoA and of carnitine palmitoyltransferase were measured in mitochondria prepared from needle biopsy samples of human skeletal muscle. Results are presented for nine normal subjects and 18 patients in whom there was evidence of mitochondrial abnormality. 2. Palmitoylcarnitine and palmitoyl-CoA oxidation were measured spectrophotometrically by following the reduction of added cytochrome c in the presence of cyanide. 3. Because of large variations in the activities between subjects it was essential to express the three activities per unit of cytochrome c oxidase activity to demonstrate unambiguous specific alterations in the activities. 4. In most of the patients the order of the three activities was similar to that in the normal subjects. However, in five cases the activity for palmitoylcarnitine oxidation was less than 4% of the mean normal value. In two of these patients, the low activity could be accounted for by very low (<10% normal) activity of carnitine palmitoyltransferase (CPT). In another two patients the activity of CPT was normal but that of palmitoyl-CoA dehydrogenase (a measure of β-oxidation) was very low.

1. An isolated skeletal muscle preparation is described which has been used to study the efflux of enzymes in response to contractile activity, metabolic poisons and detergent treatment. 2. In both fast and slow muscles contractile activity caused a release of lactate dehydrogenase and creatine kinase that reached a peak 1–2 h after the end of stimulation. There was very little release during the 30 min stimulation period whether the muscles were under aerobic or hypoxic conditions. 3. Incubation of the muscles with cyanide and iodoacetate caused a similar delayed release of enzyme. 4. Disruption of cell membranes with detergent treatment caused an entirely different and very rapid pattern of enzyme release. 5. Enzyme release from the fatigued isolated muscle preparations appears to be initiated as a consequence of phosphorylcreatine or ATP depletion. The relevance of this to release of muscle enzymes after activity in vivo is discussed.

1. A form of skeletal muscle fatigue was examined with isolated animal and human muscle preparations. The possibility that methylxanthines could overcome this was investigated. 2. Prolonged contractile activity resulted in a long-lasting impairment of force generation at low frequencies of stimulation at times when the force at higher frequencies had substantially recovered. This was seen with both fast-twitch and slow-twitch animal muscles and with samples of isolated human muscle. 3. The decrease in low-frequency force was due to a decrease in twitch amplitude, suggesting damage to the processes involved in excitation-contraction coupling. 4. Caffeine and theophylline at concentrations of 1 mmol/l rapidly and completely reversed the effects of this form of fatigue in both animal and human muscle preparations. 5. Agents that potentiate muscle force production could be an effective means of counteracting the effects of an important form of skeletal muscle fatigue, but a clinically useful compound would need to be more potent than the methylxanthines currently in use.

1. Quadriceps strength, relaxation rate, fibre-type composition and energy-turnover rate during a submaximal contraction have been measured in hypo- and hyper-thyroid patients and compared with findings in normal subjects. 2. Six out of eight hypothyroid patients had normal strength whereas four out of five hyperthyroid patients were weak. 3. Relaxation rate was decreased in all the hypothyroid patients but increased in only three out of five hyperthyroid patients. 4. In hypothyroidism there was a marked reduction in the percentage contributed by type II fibres to muscle cross-section, partly due to type II atrophy but also due to a decrease in the relative frequency of type II fibres. In hyperthyroidism both fibre types tended to atrophy. 5. The rate of ATP turnover during submaximal contraction held to fatigue was reduced in hypothyroidism. This was probably due to decreased ATP utilization rather than an impaired supply of energy-supplying substrates. In hyperthyroidism the rate of ATP turnover was increased. 6. Altered relaxation rate and ATP-turnover rate may be explained on the basis of changes in myosin ATPase activity with thyroid status. Changes in muscle-fibre-type composition, as determined histochemically, could not per se account for the functional abnormalities.

1. Muscle fibres may be subdivided into type I (with slow-twitch contractile properties) and type II (fast-twitch) depending on their myosin adenosine triphosphatase activity. In voluntary isometric contractions type I fibres are utilized at low forces (<20% of maximum) whereas type II fibres are recruited in addition at high forces. This physiological recruitment order has enabled us to measure the relaxation rate of type I and II fibres in vivo in normal human subjects. 2. Relaxation rate was measured in 16 subjects from low (10% of maximum) and maximum isometric quadriceps contractions and the muscle-fibre type composition determined from needle-biopsy specimens in 10 subjects. The relaxation rate of type II fibres was calculated to be twice as fast as that of type I. 3. It was not possible to estimate, from studies in 33 quadriceps muscles (25 normal subjects), the contribution of type II fibres to overall fibre area from the relaxation rate as determined from electrically stimulated isometric contractions.

1. To study the metabolism of α 1 -antitrypsin in man, radioiodinated purified α 1 -antitrypsin of protease inhibitor type MM was injected intravenously into seven normal volunteer subjects of protease inhibitor type MM and plasma and urine radioactivity data were analysed. The purified glycoprotein inhibited trypsin and exhibited micro-heterogeneity characteristic of α 1 -antitrypsin on acid starch-gel electrophoresis. 2. The intravascular mass was 116 ± 16.1 mg/kg (mean ± sem); extravascular/intravascular pool ratio was 1.07 ± 0.067; the fractional catabolic rate was 33.3 ± 1.37% of the intravascular pool/day; half-life was 4.6 ± 0.21 days; synthetic rate was 33.8 ± 6.3 mg day −1 kg −1 . 3. The data indicated that the serum concentration of α 1 -antitrypsin varies independently of its fractional catabolic rate and that it is catabolized in, or in close relationship to, the intravascular compartment. 4. Plasma curves of protein-bound radioactivity were also defined after the intravenous administration of 131 I-labelled desialylated α 1 -antitrypsin to two additional subjects. The fractional plasma disappearance rates were approximately 150 times as great as those for the intact protein. The rapid disappearance of 131 I-labelled desialylated α 1 -antitrypsin from plasma coincided with an equally rapid uptake of radioactivity by the liver. 5. The results are compatible with the existence of receptor sites for desialylated glycoproteins in the surface membrane of human hepatocytes and emphasize that the catabolism of α 1 -antitrypsin can be influenced by the structure of its carbohydrate moiety.

1. The fatigue of force that occurs during the first 60 s of a maximum voluntary contraction of the human quadriceps has been examined by comparing the voluntary force with that obtained by brief tetanic stimulation at 50 Hz in nine healthy subjects. In three subjects the voluntary force declined in parallel with the tetanic force whereas in the remainder it fell more rapidly, suggesting that central fatigue was present. 2. For those subjects who showed little or no central fatigue, surface electromyograph (EMG) activity remained approximately constant while the force declined by about 60%. In the others, EMG activity and force declined in parallel but when an extra effort was made the subjects could briefly increase their force and this was accompanied by a proportionately greater increase in EMG activity (generally up to the original value). 3. It is concluded that in sustained maximum voluntary contractions of the quadriceps (a) central fatigue may account for an appreciable proportion of the force loss, (b) surface EMG recordings provide no evidence that neuromuscular junction failure is the limiting factor determining the loss of force in this muscle.

1. Sixteen patients with chronic bronchitis and airways obstruction were given radioactive nitrogen ( 13 N) by intravenous injection and by inhalation, while breathing air and after 10–20 min breathing 30% oxygen. The clearance of 13 N from four zones of each patient's whole lung field was monitored. 2. The 13 N clearance of each region in these patients with chronic bronchitis was much slower than in normal subjects. Oxygen breathing produced a significant delay in the clearance of intravenously administered 13 N in 23 zones in 10 patients but no systematic change in clearance after inhaled 13 N. 3. With inhalation of 30% oxygen there was no significant change in the mean minute ventilation, tidal volume or arterial P co 2 . 4. The results suggest that local hypoxic vasoconstriction is present in some patients on breathing air and that this is relieved by 30% oxygen, resulting in a diversion of local blood flow from well-ventilated to more poorly ventilated areas. The fall in V̇ A /Q̇ on 30% oxygen is insufficient to increase arterial P co 2 .

1. The kinetics of unconjugated [ 14 C]-bilirubin metabolism have been investigated and analysed in terms of a three-pool model in a group of seven normal subjects and in a group of eight cirrhotic patients who had appreciable impairment of liver cell function. 2. In the patients with cirrhosis, the plasma unconjugated bilirubin was either normal or only slightly increased but the metabolism of unconjugated bilirubin was deranged. 3. The mean volume of distribution, the mean 4 h retention, and the mean mass of the rapidly mixing pool were all significantly greater than in the normal subjects. In contrast, mean fractional clearance rate and mean estimated erythrocyte life-span were significantly less than in the normal subjects. 4. The mean fractional transfer rates and fluxes from pool 1 (rapidly mixing pool—‘plasma’) to pool 3 (‘extravascular’) and vice versa were significantly greater than the corresponding values in the normal subjects. 5. The results indicate that, in patients with compensated cirrhosis, the efficiency of the liver in extracting unconjugated bilirubin from plasma against a concentration gradient is impaired, even though the liver's capacity to conjugate bilirubin may be normal. As a consequence of the increased volume of distribution, the absolute hepatic clearance of unconjugated bilirubin is relatively well maintained.

1. To investigate the relative degree to which human IgG and other plasma proteins bind to cell membranes of the full-term human placenta, suspensions of membranes mixed with radio-iodinated proteins were incubated at pH 6·5 and subjected to Sepharose 2B column chromatography. The amount of labelled protein associated with membrane protein in membrane-containing fractions of the eluate was then determined. 2. Binding of IgG and each of the four subclasses of IgG was appreciable. Binding of IgG was markedly reduced if membranes were incubated at pH 8·0 rather than 6·5. Binding of labelled IgG was inhibited by excess of unlabelled IgG but not by excess of unlabelled albumin. Placental membranes bound much more IgG than did erythrocyte membranes. 3. Binding of insulin was relatively greater than that of IgG, whereas binding of IgE, IgM, IgA, albumin, transferrin and polyvinylpyrrolidone was much less than that of IgG and each of its subclasses. 4. The relative binding of IgG and fragments of the IgG molecule was, in decreasing order of magnitude: light chains, IgG, Fc, heavy chains and F(ab′) 2 . 5. The results are consistent with Brambell's hypothesis for the mechanism of the transmission of passive immunity from mother to young. In particular the data are consistent with the existence of a limited number of IgG specific receptors on the microvillus membrane of the syncytial trophoblast. Such receptors could protect IgG molecules from degradation during their transplacental transport. The submolecular region of the IgG molecule involved in specific membrane binding appears to be common to all four subclasses and to include the Fc region.

1. A new method of studying isolated human skeletal muscle has been evaluated. This involves the incubation and electrical stimulation of strips of muscle, obtained at surgical biopsy, that are tied at the cut ends of the fibre bundles. 2. Morphological examination showed that the fibres were sealed off at the cut ends. Damage appeared to be restricted to the areas immediately adjacent to the ties. 3. Contractile properties were well maintained for several hours and measurements of tissue metabolites showed that muscle contents of the high-energy phosphate compounds were well preserved. 4. The isolated preparations were found to have the same contractile properties as human quadriceps femoris studied in vivo by the methods described in the preceding paper. 5. Correlation was found between the relaxation speed of the isolated preparations and their fibre-type composition histochemically determined. 6. It is concluded that this technique is a valid addition to the present methods of studying the physiology and pharmacology of human skeletal muscle.

1. The force produced by isometric contractions of the quadriceps muscle have been studied during maximal voluntary contractions and when a substantial part of the muscle was electrically stimulated via surface electrodes. 2. In normal children and adults, the force of a maximal voluntary contraction of the quadriceps was proportional to body weight. 3. The function of the quadriceps has been described in terms of the force/frequency curve, speed of relaxation and the rate of loss of force during 18 s stimulation at 30 Hz and 100 Hz. 4. The functional characteristics of adductor pollicis when stimulated via the ulnar nerve were essentially similar to those of the quadriceps. 5. Studies of the function of these two muscles are complementary since quadriceps femoris is amenable to needle biopsy investigations of its structure and chemistry whereas adductor pollicis is more suitable for electrophysiological studies.

1. Sympathetic reflexes were activated by carotid occlusion in anaesthetized dogs in which changes in renal perfusion pressure were prevented. This produced a prompt and reversible increase in plasma renin activity. 2. Administration of clonidine decreased plasma renin activity, arterial pressure and heart rate and blocked the renin secretory and blood pressure responses to carotid occlusion. 3. These results support the hypothesis that the suppression of renin secretion by clonidine is a consequence of the decrease in sympathetic activity produced by this drug.