Increased contribution of KCa channels to muscle contraction induced vascular and blood flow responses in sedentary and exercise trained ZFDM rats.

In resistance arteries, endothelium-dependent hyperpolarization (EDH)-mediated vasodilatation is depressed in diabetes. We hypothesized that downregulation of KCa channel derived EDH reduces exercise-induced vasodilatation and blood flow redistribution in diabetes. To test this hypothesis, we evaluated vascular function in response to hindlimb muscle contraction, and the contribution of KCa channels in anaesthetised ZFDM, metabolic disease rats with type 2 diabetes. We also tested whether exercise training ameliorated the vascular response. Using in vivo microangiography, the hindlimb vasculature was visualized before and after rhythmic muscle contraction (0.5 s tetanus every 3 s, 20 times) evoked by sciatic nerve stimulation (40 Hz). Femoral blood flow of the contracting hindlimb was simultaneously measured by an ultrasonic flowmeter. The contribution of KCa channels was investigated in the presence and absence of apamin and charybdotoxin. We found that vascular and blood flow responses to muscle contraction were significantly impaired at the level of small artery segments in ZFDM fa/fa rats compared to its lean control fa/+ rats. The contribution of KCa channels was also smaller in fa/fa than in fa/+ rats. Low-intensity exercise training for 12 weeks in fa/fa rats demonstrated minor changes in the vascular and blood flow response to muscle contraction. However, the KCa-derived component in the response to muscle contraction was much greater in exercise trained than in sedentary fa/fa rats. These data suggest that exercise training increases the contribution of KCa channels among endothelium-dependent vasodilatory mechanisms to maintain vascular and blood flow responses to muscle contraction in this metabolic disease rat model. KEY POINTS: Microvascular dysfunction in type 2 diabetes impairs blood flow redistribution during exercise and limits the performance of skeletal muscle and may cause early fatigability. Endothelium-dependent hyperpolarization (EDH), which mediates vasodilatation in resistance arteries, is known to be depressed in animals with diabetes. Here, we report that low-intensity exercise training in ZFDM rats increased the KCa channel-derived component in the vasodilator responses to muscle contraction compared to that in sedentary rats, partly as a result of the increase in KCNN3 expression. These results suggest that low-intensity exercise training improves blood flow redistribution in contracting skeletal muscle in metabolic disease with diabetes via upregulation of EDH.

Quadriceps Thickness and Echo Intensity Predict Gait Independence in Individuals with Severe and Mild Hemiparetic Stroke.

INTRODUCTION: Smaller muscle size and higher adipose tissue ratio of the quadriceps femoris are often observed after stroke. However, it is unclear whether muscle size and the intramuscular fat ratio of the quadriceps measured with ultrasonography (US) reflect gait independence in individuals with mild or severe hemiparetic stroke. OBJECTIVE: The present study was performed to examine the relationships of gait independence with muscle thickness (MT) and echo intensity (EI) of the quadriceps femoris in individuals with hemiparesis after stroke. METHODS: We examined 43 individuals with hemiparetic stroke. We assessed functional independence measure (FIM) gait scores and measured thickness and EI of the quadriceps using US. The relationships of FIM gait scores with MT and EI were examined using Spearman's correlation coefficients in mild (n = 21) and severe (n = 22) hemiparetic stroke groups. RESULTS: In the mild hemiparetic group, FIM gait scores were correlated with paretic limb MT (rho = 0.60, p < 0.01) and EI (rho = -0.57, p < 0.01). In the severe hemiparetic group, FIM gait scores were correlated with paretic limb MT (rho = 0.67, p < 0.01) and EI (rho = -0.43, p < 0.05), as well as non-paretic limb MT (rho = 0.86, p < 0.01) and EI (rho = -0.56, p < 0.01). CONCLUSIONS: Quadriceps thickness and EI were associated with the degree of gait independence. Atrophy and increased intramuscular fat of the quadriceps may be limiting factors for achieving gait independence.

Relationships between resting blood flow and the indices of muscle damage after eccentric contractions.

PURPOSE: The aim of the study is to examine the relationships between increments in resting blood flow and isometric maximal voluntary contraction (MVC) force reduction, muscle soreness, and swelling after eccentric contractions (ECs). METHODS: Twenty-one young healthy men (age 20.8 ± 1.6 years; height 172.0 ± 5.3 cm; weight 64.9 ± 7.7 kg) were recruited for this study. All participants performed right arm ECs in five sets of 20 repetitions with 3 min of rest between the sets. The dumbbell weight corresponded to 60% MVC force of isometric contraction of elbow flexors with 90° elbow joint angle. Resting forearm blood flow (FBF), the MVC force, the muscle thickness (MT), and muscle soreness of elbow flexors, heart rate (HR), and blood pressure (BP) of brachial artery were measured before, 24 and 48 h after ECs. RESULTS: Average and peak resting FBF after ECs significantly changed from the average values before ECs (21% and 39% increase, respectively, P < 0.01). However, resting HR and BP were not significantly different after ECs. Average increase in resting FBF showed a significant relationship with average isometric MVC force reduction (r = - 0.45, P < 0.05), peak isometric MVC force reduction (r = - 0.48, P < 0.05), average muscle soreness (r = 0.49, P < 0.05), and peak muscle soreness (r = 0.49, P < 0.05). Moreover, stepwise multiple regression analysis revealed that average increased resting FBF was explained by isometric MVC force reduction and muscle soreness (adjusted R2 = 0.33). CONCLUSIONS: These results suggested that increments in resting blood flow reflect muscle damage, and increased resting blood flow may be a result of acute inflammatory response induced by muscle damage.

Relationship of Quadriceps Muscle Thickness with Motor Paralysis and Muscle Echo Intensity in Post-Stroke Patients.

INTRODUCTION: Quadriceps muscle atrophy and quality loss, defined as an increased ratio of intramuscular fat and/or connective tissue, are often observed especially in the paretic limb of post-stroke patients. This study was performed to examine the relationship of quadriceps muscle thickness (MT) with muscle echo intensity (EI) and the severity of motor paralysis after stroke. METHODS: Thirty-six hemiparetic subacute post-stroke patients were enrolled. We examined the MT (index of muscle quantity) and the EI (index of muscle quality) at the anterior mid-thigh in both limbs. We also assessed the Brunnstrom stage (BR stage), subcutaneous adipose tissue thickness, time since stroke, age, body weight, sex, number of medications, and nutritional and inflammation status. RESULTS: The MT in the paretic limb was explained by the BR stage (ß = -0.26, p < 0.01), body weight (ß = 0.68, p < 0.01), and serum albumin (ß = 0.34, p < 0.01), with an adjusted R2 of 0.81. The MT in the non-paretic limb was explained by the muscle EI (ß = -0.55, p < 0.01) and age (ß = -0.40, p < 0.01), with an adjusted R2 of 0.69. The muscle EI was explained by the MT in the paretic limb (ß = -0.34, p < 0.01) and non-paretic limb (ß = -0.69, p < 0.01). CONCLUSIONS: Our results suggest that motor paralysis, aging, and malnutrition contribute to quadriceps atrophy in post-stroke patients. Moreover, a potential countermeasure to diminish muscle quality loss is maintenance of muscle quantity.

Relationship between trunk intramuscular adipose tissue content and prevalence of metabolic syndrome in middle-aged Japanese men.

OBJECTIVES: The aim of the present study was to examine the dose-response relationship between trunk tissue composition and prevalence of metabolic syndrome (MetS) in middle-aged Japanese men. METHODS: The 1026 men (between 35 and 59 y of age) who participated in the present study were divided into two groups: those with metabolic syndrome (MetS) and those without (non-MetS). Intramuscular adipose tissue (IntraMAT) content and the cross-sectional areas (CSAs) of visceral adipose tissue and skeletal muscle tissue were calculated using low-dose computed tomography images acquired at the level of the third lumbar vertebra. Height, body mass, body fat, waist circumference, the presence of MetS, and lifestyle habits were also assessed. RESULTS: IntraMAT content was significantly higher in MetS than in non-MetS men. A 10% increase in IntraMAT content correlated with the prevalence of MetS (odds ratio, 4.197; 95% confidence interval, 3.108-7.088; P < 0.001), even after adjustments for age, height, adjusted skeletal muscle CSA, sleeping time, alcohol consumption, exercise habit, and cigarette smoking. Skeletal muscle CSA did not correlate with the prevalence of MetS after adjustments for IntraMAT content and other cofactors. CONCLUSIONS: Increase in IntraMAT content, not in skeletal muscle CSA, significantly correlated with the prevalence of MetS. These results suggest that countermeasures against the accumulation of trunk IntraMAT effectively prevent MetS in middle-aged Japanese men.

Skeletal muscle and abdominal circumference explain intramuscular fat, independent of exercise frequency, in middle-aged Japanese men.

The purpose of this study was to examine how intramuscular adipose tissue (IntraMAT) can be characterized using physical and functional characteristics, muscle size, and/or adipose tissue in four different exercise frequency groups of middle-aged Japanese men. One thousand twenty-eight middle-aged men (age, 47.5 ± 8.1 years; height, 170.5 ± 5.8 cm; body mass, 67.0 ± 9.3 kg; body mass index, 21.9 ± 5.3 kg/m2) were allocated to four groups on the basis of their weekly exercise frequency: Group 1, no exercise, n = 334; Group 2, once a week, n = 271; Group 3, a few days a week, n = 269; and Group 4, every day, n = 154. Their body composition, blood pressure, and handgrip strength were assessed. A single-slice computerized tomography image at the level of the umbilicus was acquired and the CSAs of IntraMAT, muscle tissue, and subcutaneous and visceral adipose tissues (SCAT and VAT) were calculated. The %IntraMAT significantly correlated with physical characteristics, such as age, BMI, abdominal circumference, and muscle tissue CSA. Stepwise multiple regression analysis was performed, with the %IntraMAT as the dependent variable. Muscle tissue CSA and abdominal circumference were the common independent variables across groups to explain the variability of the %IntraMAT. It was also extracted %body fat and age for Group 2, age, handgrip strength, and BMI for Group 3, and smoking category for Group 4. These results suggested that muscle tissue size and abdominal circumference would be the strong predictors to explain %IntraMAT of the trunk muscle across four groups, and that age, %body fat, BMI, and SCAT, handgrip strength and smoking category were also good predictors for each group with different exercise frequency in middle-aged Japanese men.

Contribution of skeletal muscle and serum lipids to muscle contraction induced by neuromuscular electrical stimulation in older individuals.

Intramyocellular lipids (IMCL) stored in droplets in muscle cells and free fatty acids (FFA) from fat cells in the blood are the main substrates of adenosine triphosphate during continuous muscle contractions of relatively lower intensity. Although it is known that the lipid oxidative capacity decreases with aging, the effect of IMCL and FFA on muscle contraction in older individuals remains unclear. The purpose of this study was to investigate the contribution of skeletal muscle lipids and blood lipids as energy sources for muscle contraction in older individuals. Eighteen older individuals (mean age: 70.4 ± 3.5 years) underwent muscle contraction intervention induced by intermittent neuromuscular electrical stimulation (NMES) to the vastus lateralis for 30 min. Fasting blood samples were obtained and proton magnetic resonance spectroscopy (1 H-MRS) was performed before and after NMES, and the parameters (including IMCL and extramyocellular lipid [EMCL]) from 1 H-MRS, along with FFA and adiponectin levels, were analyzed using the blood samples of all participants. Levels of IMCL and EMCL did not change (p > 0.05); however, FFA and adiponectin levels decreased from 1.1 ± 0.5 mEq/L to 0.8 ± 0.2 mEq/L and 12.0 ± 5.3 µg/ml to 11.4 ± 5.0 µg/ml, after NMES (p < 0.05), respectively. These findings indicate that serum lipids, but not skeletal muscle lipids, are the energy substrate utilized during involuntary muscle contraction in older individuals.

Importance of skeletal muscle lipid levels for muscle function and physical function in older individuals.

The skeletal muscle contains lipids inside (intramyocellular lipids, IMCL) or outside (extramyocellular lipids, EMCL) its cells. The muscle lipid content increases with age; however, the characteristics of IMCL and EMCL in older individuals are not well known. We aimed to examine the characteristics of skeletal muscle lipids by investigating their relationship with muscle function and physical functions. Seven elderly men and 16 elderly women participated. The skeletal muscle lipid content, including IMCL and EMCL, was measured in the vastus lateralis by proton magnetic resonance spectroscopy. Isometric knee extension with maximal voluntary contraction (MVC) and time-to-task failure for knee extension with 50% MVC were measured as muscle functions. The participants performed six physical function tests: preferred gait speed, maximal gait speed, Timed Up and Go, chair sit-to-stand, handgrip strength, and stand from the floor. The time to knee extension task failure had a significant relationship with the IMCL (rs = -0.43, P < 0.05), but not with the EMCL content. Significant relationships were confirmed in the EMCL content with the sit-to-stand (rs = -0.48, P < 0.05) and stand-from-the-floor (rs = 0.53, P < 0.05) tests. These findings indicated that muscle lipids are associated with muscle and physical functional performances in older individuals. Novelty: No relationship was confirmed between IMCL and EMCL in older individuals. Muscle endurance performance had a relationship with IMCL, but not with EMCL. Relationships between EMCL and physical functional tests (e.g., sit-to-stand and stand from the floor) were confirmed.

Effect of elbow joint angles on electromyographic activity versus force relationships of synergistic muscles of the triceps brachii.

The electromyographic (EMG) activity and force relationship, i.e. EMG-force relationship, is a valuable indicator of the degree of the neuromuscular activation during isometric force production. However, there is minimal information available regarding the EMG-force relationship of individual triceps brachii (TB) muscles at different elbow joint angles. This study aimed to compare the EMG-force relationships of the medial (TB-Med), lateral (TB-Lat), and long heads (TB-Long) of the TB. 7 men and 10 women performed force matching isometric tasks at 20%, 40%, 60%, and 80%maximum voluntary contraction (MVC) at 60°, 90°, and 120° of extension. During the submaximal force matching tasks, the surface EMG signals of the TB-Med, TB-Lat, and TB-Long were recorded and calculated the root mean square (RMS). RMS of each force level were then normalized by RMS at 100%MVC. For the TB-Med, ultrasonography was used to determine the superficial region of the muscle that faced the skin surface to minimize cross-talk. The joint angle was monitored using an electrogoniometer. The elbow extension force, elbow joint angle, and surface EMG signals were simultaneously sampled at 2 kHz and stored on a personal computer. The RMS did not significantly differ between the three muscles, except between the TB-Med and TB-Lat during 20%MVC at 60°. The RMS during force levels of ≥ 60%MVC at 120° was significantly lower than that at 60° or 90° for each muscle. The sum of difference, which represents the difference in RMS from the identical line, did not significantly differ in any of the assessed muscles in the present study. This suggests that a relatively smaller neuromuscular activation could be required when the elbow joint angle was extended. However, neuromuscular activation levels and relative force levels were matched in all three TB synergists when the elbow joint angle was at 90° or a more flexed position.

Comparison of metabolic risk factors, physical performances, and prevalence of low back pain among categories determined by visceral adipose tissue and trunk skeletal muscle mass in middle-aged men.

The present study compared metabolic risk factors, physical performances, and musculoskeletal impairment among categories determined by visceral adipose tissue (VAT) and trunk skeletal muscle (SM) mass in middle-aged Japanese men. In total, 1026 healthy Japanese males aged between 35 and 59 years were categorized into 4 groups according to the amount of VAT and SM in the trunk measured using low-dose computed tomography (LowVAT-HighSM, LowVAT-LowSM, HighVAT-HighSM, and HighVAT-LowSM). Height, body mass waist circumference, body fat, intramuscular adipose tissue (IntraMAT), subcutaneous adipose tissue, biochemical blood profiles (triglycerides, high-density lipoprotein cholesterol, fasting blood glucose, aspartate transaminase, alanine transaminase and γ-glutamyl trans peptidase), physical performances (trunk flexibility, the chair-stand test, two-step length and hand-grip strength), the prevalence of low back pain, and lifestyle habits for exercise, alcohol intake and smoking, were compared among the groups. The results showed that LowVAT-HighSM had significantly superior biochemical blood profiles and physical performances to the other groups. HighVAT-LowSM had significantly higher %IntraMAT and the prevalence of low back pain. The two-step length, which is an index of walking ability, significantly differed according to the four subject categories. These results indicate that metabolic risk factors, physical performances, and prevalence of low back pain in middle-aged Japanese men may differ among four categories determined by VAT and trunk SM.

Chair sit-to-stand performance is associated with diagnostic features of sarcopenia in older men and women.

BACKGROUND: Standing from a chair is a fundamental activity of daily living, and it can be applied to assess the physical function, especially in older individuals. AIM: The aim of this study was to elucidate the characteristics of mechanical and temporal parameters during chair stand based on the relationship with skeletal muscle and physical functional parameters in older men and women. METHODS: Eighty older men and women participated in this study. We measured four parameters of chair stand performance: ground reaction force (GRF), rate of force development (RFD), and chair rise time (CRT) were calculated from the foot-floor force data; sit-to-stand (STS) was also assessed by measuring the time needed to complete 10 chair stand repetitions. The muscle thickness (MT) and echo intensity, as indexes of muscle size and quality, respectively, were measured using axial B-mode ultrasound images from quadriceps femoris. The gait speed and handgrip strength were measured as physical functional parameters. RESULTS: Partial correlation was used to determine the association of chair stand performance with MT, echo intensity, and physical parameters while considering the height, body mass, and age. GRF, RFD, and STS were significantly correlated with MT (r = 0.35, 0.26, and -0.49), gait speed (r = 0.32, 0.31, and -0.67), and handgrip strength (r = 0.57, 0.59, and -0.49). As the result of regression analysis, MT, gait speed, and handgrip strength were estimated by GRF and STS. CONCLUSION: These results suggest that chair stand performance is useful as it reflects the muscle size and physical functions in older individuals.

Comparison of fascicle behaviors between superficial and deeper muscles of triceps brachii during isometric contractions.

PURPOSE: We assessed fascicle behaviors of the upper extremities during isometric contractions at different joint angles in this study. METHODS: Thirteen healthy men and women performed isometric elbow extension tasks at 50% and 75% of maximal voluntary contraction (MVC) at 60°, 90°, and 120° of elbow extension (full extension = 180°). Extended field-of-view B-mode ultrasonography was used to obtain sagittal plane panoramic images of the long head (TB-Long) and medial head (TB-Med) of the triceps brachii at rest and during contraction; fascicle length and pennation angle were measured. RESULTS: In the TB-Long, significant fascicle shortening from rest was found during 50% and 75%MVC at 60° and during 75%MVC at 90° of extension. There was no significant fascicle shortening in the TB-Med muscle under any conditions. There was no significant pennation angle change from rest in either muscle. The pennation angle of the TB-Long was significantly greater than that of the TB-Med under all conditions. CONCLUSIONS: These results suggest that fascicle shortening in the TB-Long muscle occurs in flexion; however, no change was found in the TB-Med. In the upper extremity muscle-tendon complex, the superficial and deeper muscles may have different force-transmission efficiency at flexed joint angles.

Quadriceps echo intensity can be an index of muscle size regardless of age in 65 or more years old.

Although age-related deterioration of skeletal muscle strength and size has gained much attention, the age-related increase in adipose tissue within skeletal muscle has not received the same level of attention, especially in older individuals. It is not well known how echo intensity (EI), which is an index of intramuscular adipose tissue (IntraMAT) and connective tissue, is associated with demographic, functional, and morphological characteristics in older individuals. The purpose of this study was to assess the relationships between the EI of the quadriceps femoris (QF) and demographic, functional, and morphological characteristics in pre-old (65-74 years) and old (75 years or older) individuals. One hundred thirty-two men and women aged 65 to 74 years (45 men and 51 women; Pre-old group) or 75 years and older (17 men and 19 women, Old group) participated in this study. The EI and muscle thickness (MT) of the QF were measured using B-mode ultrasonography. Sit-up, supine-up, sit-to-stand, 5-m maximal walk, and 6-min walk tests were performed. The EI of the QF in the Old group (73.1 ± 9.0 a.u.) was significantly higher than that in the Pre-old group (67.6 ± 9.1 a.u.). The EI of the QF was significantly correlated with the MT of the QF in both the Pre-old (r = -0.542, P < 0.01) and Old (r = -0.622, P < 0.01) groups. Stepwise multiple linear regression analysis with the EI of the QF as a dependent variable showed that the MT of the QF was a significant variable in both the Pre-old and Old groups. These results suggest that MT is an extremely valuable factor for explaining the EI of the QF regardless of age group. Exercise interventions are recommended to decrease the risks of metabolic impairment by increasing muscle size and decreasing IntraMAT in individuals aged 65 years or older.

Comparison of muscle quality and functional capacity between Japanese and Brazilian older individuals.

Muscle quality is well-known to decrease with aging and is a risk factor for metabolic abnormalities. However, there is a lack of information on race-associated differences in muscle quality and other neuromuscular features related to functional performance. This study aimed to compare muscle quality, function, and morphological characteristics in Japanese and Brazilian older individuals. Eighty-four participants aged 65-87 years were enrolled in the study (42 Japanese: 23 men, 19 women, mean age 70.4 years; 42 Brazilians: 23 men, 19 women, mean age 70.8 years). Echo intensity (EI) and muscle thickness (MT) of the quadriceps femoris were measured using B-mode ultrasonography. A stepwise multiple linear regression analysis with EI as a dependent variable revealed that MT was a significant variable for Japanese participants (R2 = 0.424, P = 0.001), while MT and subcutaneous adipose tissue (SCAT) thickness were significant variables for Brazilian participants (R2 = 0.490, P = 0.001). A second stepwise multiple linear regression analysis was performed after excluding MT and SCAT thickness from the independent variables. Sex and age for Japanese participants (R2 = 0.381, P = 0.001) and lean body mass and body mass index for Brazilian participants (R2 = 0.385, P = 0.001) were identified as significant independent variables. The present results suggest that MT is closely correlated with muscle quality in Japanese and Brazilian older individuals. Increases in muscle size may induce decreases in intramuscular adipose tissue and/or connective tissues, which are beneficial for reducing the risks of metabolic impairments in Japanese and Brazilian older individuals.

Regulation of the microvascular circulation in the leg muscles, pancreas and small intestine in rats.

To study the microvascular circulation, we examined the proportion of open and functioning capillaries in the leg muscles, pancreas and small intestine of anesthetized rats. Fluorescein isothiocyanate (FITC)-labeled Lycopersicon esculentum lectin was injected into the heart and allowed to circulate for 3 min to label open and functioning capillaries. Specimens were removed, frozen, sectioned and double-immunostained. Using one section, open and functioning capillaries were detected by immunostaining for this lectin bound to endothelial cells, while all capillaries were visualized by immunostaining for platelet endothelial cell adhesion molecule-1 (PECAM-1 or CD31). These capillaries were semi-automatically detected and counted by fluorescence microscopy. The percentages of open and functioning capillaries were as follows: the soleus muscle, 93.0 ± 5.5%; superficial zone of the gastrocnemius muscle, 90.8 ± 6.2%; deep zone of the gastrocnemius muscle, 95.6 ± 4.0%; the plantaris muscle, 94.1 ± 2.7%; the pancreas, 86.3 ± 11.7%; and the small intestine, 91.1 ± 4.9% (n = 8, each). There was no significant difference among these data by the Kruskal-Wallis test. This study clearly demonstrated that the proportions of open and functioning capillaries are high and similar among the leg muscles, pancreas and small intestine in spite of their structural and functional differences. This finding agrees with previous studies and supports the notion that the microvascular circulation is mainly controlled by changing of the blood flow in each capillary rather than changing the proportion of open and functioning capillaries.

Microvascular circulation at cool, normal and warm temperatures in rat leg muscles examined by histochemistry using Lycopersicon esculentum lectin.

Local cooling and/or warming of the body are widely used for therapy. For safer and more effective therapy, microvascular hemodynamics needs to be clarified. To examine blood circulation in rat leg muscles at 20, 30, 37 and 40°C, fluorescein isothiocyanate (FITC)-labeled Lycopersicon esculentum lectin was injected into the cardiac ventricle. Endothelial cells of open and functioning blood vessels were labeled by this lectin for 3 min and detected by immunostaining for lectin. The percentage of open and functioning capillaries of leg muscles by the avidin-biotin method was 89.8±3.3% at 37°C, while capillaries were unclear or unstained at 20 and 30°C, probably due to a decrease of blood flow. The results using the tyramide-dinitrophenol method were 58.6±15.0% at 20°C, 68.5±12.3% at 30°C, 83.8±5.7% at 37°C and 83.3±7.8% at 40°C. The value at 20°C was significantly different from those at 37 and 40°C. The results by the tyramide-biotin method were 85.5±5.3% at 20°C, 87.3±9.7% at 30°C, 94.7±3.6% at 37°C and 92.5±2.1% at 40°C. Based on these results, it was concluded that the blood flow of each capillary considerably decreased at 20 and 30°C and probably increased at 40°C, whereas the proportion of open and functioning capillaries was essentially unchanged.