Preliminary Communication: Familial Accumulation of Carriers of Au Antigen

The sibs and parents of 83 Au-positive blood donors were tested in order to study the possible family clustering of Au positives in a population with a low frequency of carriers of Australia (Au) antigen. Tested by counterelectrophoresis the incidence of Au antigen in the parents of Au-positive persons is 5·4% and in the sibs 17·9%, compared to 0·15% in blood donors.

The hypothesis that the Au-carrier state is genetically determined was tested by calculating the proportion of Au-positive children in the families. The results are not inconsistent with a recessive mode of inheritance but do not allow definitive conclusions to be drawn.

     

Segregation Analysis of A Large t(21q22q) Family

A large family in which a t(21q22q) chromosome is segregating is described. Segregation analysis using data from this family and previously published families shows the risk to a female carrier of having a child with translocation Down's syndrome to be 0·0684±0·0270. The risk to a male carrier is probably much smaller with an upper limit of 0·0286. The segregation ratio for phenotypically normal offspring of carrier parents of both sexes is 0·5053±0·0524.     

Effect of Anteromedial Portal Entrance Drilling Angle during Anterior Cruciate Ligament Reconstruction: A Three-Dimensional Computer Simulation

Purpose

The object of this study was to evaluate entrance angle effects on femoral tunnel length and cartilage damage during anteromedial portal drilling using three-dimensional computer simulation.

Materials and Methods

Data was obtained from an anatomic study performed using 16 cadaveric knees. The anterior cruciate ligament femoral insertion was dissected and the knees were scanned by computer tomography. Tunnels with different of three-dimensional entrance angles were identified using a computer simulation. The effects of different entrance angles on the femoral tunnel length and medial femoral cartilage damage were evaluated. Specifically, tunnel length and distance from the medial femoral condyle to a virtual cylinder of the femoral tunnel were measured.

Results

In tunnels drilled at a coronal angle of 45°, an axial angle of 45°, and a sagittal angle of 45°, the mean femoral tunnel length was 39.5±3.7 mm and the distance between the virtual cylinder of the femoral tunnel and the medial femoral condyle was 9.4±2.6 mm. The tunnel length at a coronal angle of 30°, an axial angle of 60°, and a sagittal angle of 45°, was 34.0±2.9 mm and the distance between the virtual cylinder of the tunnel and the medial femoral condyle was 0.7±1.3 mm, which was significantly shorter than the standard angle (p<0.001).

Conclusion

Extremely low and high entrance angles in both of axial plane and coronal plane produced inappropriate tunnel angles, lengths and higher incidence of cartilage damage. We recommend that angles in proximity to standard angles be chosen during femoral tunnel drilling through the anteromedial portal.     

Quantitative aspects of sensitivity and summation in the cat retina

1. Properties of the central response mechanism of on-centre ganglion cells in the cat retina were studied by recording, from optic tract fibres, responses evoked by stimuli modulated with time in a sinusoidal or square-wave fashion.

2. The shape of averaged square-wave responses resulting from the central mechanism alone was identified. This shape was identical from one cell to another. Such an identification permits the early recognition of peripheral antagonism.

3. Threshold sensitivity for a sinusoidal stimulus was determined for fifty cells along one horizontal and vertical axis, passing through the most sensitive portion of the receptive field. These sensitivity profiles were described in terms of a central segment of constant maximum sensitivity (uniform centre) and sloping outer segments of exponentially decreasing sensitivity (exponential annulus). The dimensions of the uniform centre (horizontal axis × vertical axis) varied from 0·1° × 0·1° to 2·5° × 2·2°, the half width of the exponential annulus ranged from 0·1° to 0·63°.

4. Adapting spots of varying diameter were placed concentric with the receptive field and the (unmodulated) luminance, at each diameter, that reduced a small central (sinusoidal) stimulus to threshold, was determined. The resulting area—adaptation curve, (adapting luminance plotted against diameter) showed that within defined limits the state of adaptation is determined by the flux independent of its distribution.

5. Sinusoidal stimuli of varying diameter were placed concentric with the receptive field and the threshold luminance at each diameter was determined. Suprathreshold square-wave stimuli indicated that the central mechanism alone contributed to the response. These area-sensitivity curves did not show any decrease in sensitivity at larger diameters.

6. The shape of the area—sensitivity curve, and hence the extent of the summating area, was found to be independent of the state of adaptation.

7. For any one cell the shapes of the area—adaptation and area—sensitivity curves were shown to be identical, indicating that adapting flux and stimulus flux are independent of distribution over the same defined limits.

8. The sensitivity of combinations of small disconnected areas of the receptive field was found to be equal to the sum of their individual sensitivities.

     

Genetics of plasma paroxonase activity

Complex segregation analysis of plasma paroxonase activity gives evidence for a major locus with incomplete dominance (d = 0·70) causing high activity plus a background of multifactorial inheritance (heritability = 0·09).     

Determination of three-dimensional muscle architectures: validation of the DTI-based fiber tractography method by manual digitization

In the last decade, diffusion tensor imaging (DTI) has been used increasingly to investigate three-dimensional (3D) muscle architectures. So far there is no study that has proved the validity of this method to determine fascicle lengths and pennation angles within a whole muscle. To verify the DTI method, fascicle lengths of m. soleus as well as their pennation angles have been measured using two different methods. First, the 3D muscle architecture was analyzed in vivo applying the DTI method with subsequent deterministic fiber tractography. In a second step, the muscle architecture of the same muscle was analyzed using a standard manual digitization system (MicroScribe MLX). Comparing both methods, we found differences for the median pennation angles (P < 0.001) but not for the median fascicle lengths (P = 0.216). Despite the statistical results, we conclude that the DTI method is appropriate to determine the global fiber orientation. The difference in median pennation angles determined with both methods is only about 1.2° (median pennation angle of MicroScribe: 9.7°; DTI: 8.5°) and probably has no practical relevance for muscle simulation studies. Determining fascicle lengths requires additional restriction and further development of the DTI method.     

Immune systems of human nasopharyngeal and palatine tonsils: histomorphometry of lymphoid components and quantification of immunoglobulin-producing cells in health and disease

The relative contributions of the various lymphoid tissue compartments (follicle centre, mantle zone, extrafollicular area, and reticular epithelium) of clinically normal nasopharyngeal (adenoid) and palatine tonsils of young children (mean age 5, range 2–10 years) were found to be almost identical in the two organs. In adenoid hypertrophy and recurrent palatine tonsillitis a significant relative reduction of the contributions of follicle centres and mantle zones occurred along with an increase of the extrafollicular compartment. The tonsillar Ig-producing cell systems in health and disease were studied by paired immunohistochemical staining. Only occasional IgE-producing cells were seen in normal as well as in diseased specimens. A general predominance of IgG cells followed by the IgA, IgM and IgD classes was demonstrated in both organs. In the palatine tonsils the percentage class ratios (64·4:29·6:4·3:1·7) were virtually unaltered in disease. Conversely, the class ratios were changed from 54·2:35·2:7·4:3·3 in healthy adenoids to 70·3:23·3:4·7:1·7 in hypertrophy. Intensified B-cell stimulation associated with disease was indicated as the follicle centres tended to show an increased density of Ig-producing cells—at least in the palatine tonsils—and, moreover, showed a significant shift to IgG expression. Taking the histomorphometric data into account, a decreased number of extrafollicular Ig-producing cells per tissue unit was indicated in recurrent tonsillitis. This change was most likely secondary to the disease. Conversely, the hypertrophic adenoids showed significantly increased B-cell activity in the extrafollicular compartment—a raised number of IgG-producing cells being especially marked. A difference between the two organs in the response pattern of the Ig-producing cells, especially in the reticular epithelium, might be influenced by a contribution of the secretory immune system to the surface protection of the adenoids.     

Anti-thyroid drugs and lymphocyte function: II. The in vivo effect on blastogenesis and suppressor cell activity in graves' disease

The proliferative response (PR) of T lymphocytes in PHA stimulated cultures (5 μg/ml and 0·5 μg/ml; 72 hr) was used to assess the suppressive capabilities of peripheral blood mononuclear cells (PBMC) in the thyrotoxic phase of Graves' disease and their possible modification by propylthiouracil (PTU) and methimazole (MMI) treatment. Graves' patients had 50% higher PR than controls. Treatment with PTU (n=6) for 9·5 weeks (mean) or MMI (n=6) for 18 weeks (mean) resulted in continuous decrease in PR, starting after 3 weeks and down to control values and plateau at 7-10 weeks. This decrease correlated with the decline in plasma thyroxine (T4) levels which had already dropped by 3 weeks. Grouped according to thyroid functional state PR was significantly decreased only in the euthyroid state. Suppressor cell function, expressed as suppressor removal index (PR of PBMC pre-incubated for 24 hr/PR of fresh PBMC), was significantly lower in Graves' patients compared to controls and reached above normal values under PTU treatment: 0·98±0·16, 1·39±0·09 and 1·94±0·19 (mean±s.e.m.) respectively. A direct suppressive effect of PTU in culture, observed in normal subjects, did not exist in untreated patients and evolved under MMI treatment to above normal levels. The cell-mediated PTU effect, exercised by PBMC pre-incubated with PTU on autologous cells pre-incubated in medium alone, increased under PTU treatment to above normal levels. Both this cell-mediated suppressive effect and augmented PR of pre-incubated cells were already significantly increased after 3 weeks of PTU treatment, when all patients were still thyrotoxic. We conclude therefore that PBMC of patients in the untreated, thyrotoxic phase of Graves' disease are deficient in an active cell-mediated suppressive function, a deficiency corrected—with compensatory overshoot—during anti-thyroid drug treatment.     

Accuracy of 2D CT-Based Measurements of Rotational Alignment of the Femoral Component for Total Knee Arthroplasty

PurposeWe aimed to investigate the accuracy of two-dimensional computed tomography (2D-CT)-based methods for measuring rotational alignment of the femoral component during total knee arthroplasty in comparison to reference values for three-dimensional (3D) reconstruction.Materials and MethodsWe selected the “most protruding transepicondylar axis section,” “most protruding posterior condylar line section,” and “distal femoral cut section” on 2D-CT images for 100 knees. We investigated posterior condylar angle (PCA) and condylar twist angle (CTA) values using three different methods on 2D-CT and compared to these values to those obtained using a 3D model.ResultsThe mean PCA and CTA values were 2.8° and 7.0° on the 3D model and 2.0° to 2.1° and 5.9° to 6.0° on 2D-CT, respectively. Errors in PCA and CTA measurement included internal rotation of 0.8° and 1.1° with the 1-plane and 2-plane methods and 0.9° and 1.0° with the assumed resection method, respectively.ConclusionMean errors in PCA and CTA values measured using three different methods on 2D-CT were not significantly different. However, PCA and CTA values measured on 2D-CT were approximately 1° smaller than their 3D values. Thus, we suggest that adding 1° to the mean PCA and CTA values obtained from a single plane of 2D-CT would provide values similar to those obtained from 3D reconstruction.     

A study of the mechanism of water transfer across frog skin by a comparison of the permeability of the skin to deuterated and tritiated water

1. Frog abdominal skins were placed between the two half-cells of a modified version of Ussing's frog skin apparatus.

2. The rate of equilibration of deuterated and tritiated water across the skin was followed at pH 5·0, 6·9, and 8·0 with Ringer solution bathing both surfaces of the skin; and at pH 7·0 with sulphate Ringer solution bathing both surfaces of the skin.

3. The ratio of the permeability coefficients for the influx of deuterated and tritiated water had a mean value over all the experiments of 0·9954 ± 0·0207, n = 27, indicating that no isotope effect was observable within the limits of accuracy of the experimental method.

4. The thickness of the unstirred layers associated with the frog skin was measured, and, from the data obtained, values of the true permeability coefficients for each isotope were calculated.

     

How to Minimize Rotational Conflict between Femoral & Tibial Component in Total Knee Arthroplasty: The Use of Femoro-Tibial Axial Synchronizer (Linker)

PurposeThe purpose of this study was to investigate the correlation between rotational axes of femur and tibia with the use of Linker.ResultsThe tibial component was internally rotated for the femoral component at an angle of 0.8°. The femoral component was externally rotated for the surgical transepicondylar axis (TEA) at an angle of 1.6 (range: from 4.8° of internal rotation to 7.9° of external rotation, SD=2.2°), and the tibial component was externally rotated for the surgical TEA at an average angle of 0.9 (range: from 5.1° of internal rotation to 8.3° of external rotation, SD=3.1°).ConclusionThe femoro-tibial synchronizer helped to improve the orientation and positioning of both femoral component and tibial component, and also increase the correlation of the rotational axes of the two components.     

The functional polymorphisms of VDR,GC and CYP2R1 are involved in the pathogenesis of autoimmune thyroid diseases

Vitamin D is a multi-functional immune regulator, and a low serum concentration of vitamin D promotes autoimmune inflammation. In this study, we evaluate the association between the prognosis of autoimmune thyroid disease (AITD) and the functional polymorphisms of genes that regulate vitamin D metabolism. For 139 Graves’ disease (GD) patients, 116 Hashimoto''s disease (HD) patients and 76 control subjects, we genotyped the following polymorphisms using polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP): vitamin D receptor (VDR): rs731236, rs7975232, rs2228570 and rs1544410; group-specific component (GC): rs7041 and rs4588; and CYP2R1: rs10741657. The frequency of the TT genotype for the rs731236 polymorphism was higher in GD patients than in HD patients (P = 0·0147). The frequency of the C allele for the rs7975232 polymorphism was higher in GD patients than in control subjects (P = 0·0349). The proportion of GD patients whose anti-thyrotrophin receptor antibody (TRAb) level was >51% was higher in those with the CC genotype than in those with the CA+AA genotypes (P = 0·0065). The frequency of the CC genotype for the rs2228570 polymorphism was higher in HD patients than in control subjects (P = 0·0174) and GD patients (P = 0·0149). The frequency of the Gc1Gc1 genotype for the GC polymorphism and the AG genotype for the CYP2R1 polymorphism were lower in intractable GD than in GD in remission (P = 0·0093 and 0·0268, respectively). In conclusion, genetic differences in the VDR gene may be involved in the development of AITD and the activity of GD, whereas the genetic differences in the GC and CYP2R1 genes may be involved with the intractability of GD.     

Frequency of deletion of short arm satellites in acrocentric chromosomes

The frequency of deletion of short arm satellites has been examined in various populations. Four out of 2509 males in psychiatric institutions and prisons had Dps- or Gps- (1·6 per 1000), and five out of 6187 persons in the general population had Dps- or Gps- (0·8 per 1000). The difference is not significant.

The segregation rate of the chromosome with lack of short arm satellite material did not deviate from unity in the few families where such calculations were possible. There was no increase in abortions or non-disjunction in the families with an acrocentric chromosome lacking short arm satellite material.

The study indicates that the frequency of Dps- and Gps- in the population is between 0·5 and 1·0 per 1000 and that the lack of short arm satellite material is without any deleterious effect on phenotype.

     

Increased numbers of CD5+ B cells and T cell receptor (TCR) γδ+ T cells are associated with younger age of onset in rheumatoid arthritis (RA)

Patients presenting with RA before the age of 45 years (younger onset) are known to have more aggressive disease compared with patients presenting after the age of 65 years (older onset). Coordinated expansion of circulating CD5⁺ B cell and TCR γδ⁺ T cell levels has been reported in patients with RA. This study assesses the peripheral blood levels of these two cell types in RA patients with younger and older onset of disease. CD5⁺ B cell levels were significantly elevated in the younger onset RA group (26·6 ± 4·5%) compared with the older onset RA group (14·2 ± 1·2%; P <0·01). TCR γδ⁺ T cell levels were also significantly raised in the young patients (4·0 ± 0·9%) compared with elderly patients (1·6 ± 0·2%; P <0·01). T cell levels (CD3⁺) were similar in both groups (young 66·4 ± 3·3%; old 74·3 ± 3·4% (mean ± s.e.m.); NS). Total B cell levels (CD19⁺) were also similar in these groups (7·7 ± 0·7% versus 8·9 ± 1·8%; NS). A significant positive correlation was observed between the CD5⁻ B and TCR γδ⁺ T cell types in the patients (r = 0·72, P <0.05). Compared with age-matched normal controls, the younger onset patients had similar CD5⁺ B cell and TCR γδ⁺ T cell levels to the elderly controls (CD5⁺ B cells 30·2 ± 3·0%; TCR γδ⁺ T cells 3·0 ± 0·8%). Conversely, older onset RA patients had CD5⁺ B cell levels similar to the young controls (12·3 ± 1·9%). Spontaneous in vitro synthesis of immunoglobulins (IgM, IgA and IgG) and rheumatoid factors (IgM and IgA isotypes) were not significantly different in both patient groups. The coordinate expansion of circulating CD5⁺ B cells and γδ⁺ T cells seen in patients with RA presenting before 45 years of age and not after 65 years of age may suggest a potential role for these cells in more aggressive disease states.     

Validation of digit-length ratio (2D:4D) assessments on the basis of DXA-derived hand scans

Background

The second-to-fourth digit-length ratio (2D:4D) may be a correlate of prenatal sex steroids, and it has been linked to sporting prowess. The aim of the study was to validate dual-energy X-ray-absorptiometry (DXA) as a technique to assess 2D:4D in soccer players under 15 years of age (U-15).

Methods

Paired X-ray and DXA scans of the left hands of 63 male U-15 elite soccer players (age: 14.0 ± 0.3 years) were performed, and 2D:4D was then compared between the two techniques. The 2D:4D measurements were performed twice by two blinded raters. Intrarater and interrater reliability, as well as agreement between the X-ray and the DXA assessments, were tested.

Results

Intrarater reliabilities of both raters using X-ray with intraclass correlation coefficients (ICCs) of 0.97 and 0.90 were excellent. Using DXA, the ICCs were 0.90 and 0.91 thus also showing excellent reliability. Interrater reliabilities were excellent using both the X-ray (ICC of 0.94) and the DXA (ICC of 0.90), assessments respectively. Bland-Altman plots demonstrated that the 2D:4D ratios of the two raters did not differ significantly between the X-ray and the DXA assessments. The standard errors of estimate were 0.01 for both techniques. The 95% limits of agreement of ±0.018 (±2.0%) and ±0.023 (±2.6%), respectively, were within the acceptable tolerance of 5%, and showed very good agreement.

Conclusion

DXA offered a replicable technique for assessing 2D:4D in youth soccer players. Therefore, the DXA technique seems to be an alternative method for evaluating 2D:4D in youth sports.     

Refractive error among urban preschool children in Xuzhou,China

The prevalence of refractive errors in urban preschool children in Xuzhou, China remains unknown. Children attending twelve randomly selected kindergartens participated in this study. Visual acuity, ocular alignment, cover-uncover test, cycloplegic refraction, slit-lamp and funduscopy were performed under a standardized testing environment. Cycloplegic streak retinoscopy was performed for all subjects. The mean spherical equivalent (SE) refractive error was the main outcome measure. Emmetropia was defined as refractive status between +1.75 diopters (D) and -0.75D. Myopia, hyperopia, astigmatism and anisometropia were defined as SE < -0.50D, SE > +2.0 D, cylindrical error > 1.0 D and SE difference ≥ 1 D between fellow eyes, respectively. Out of 2349 eligible children, 2255 (96%) children completed a refractive examination. Of the 2255 children, the mean SE of right eyes was +1.14 ± 0.95 diopters (D). Mean SE of the right eyes did not decline with age (r = -0.01; P = 0.56). The majority (86.6%) of children were emmetropia. The prevalence of myopia and hyperopia was 0.9% and 14.3%, respectively. The mean astigmatism for the right eyes was 0.87 ± 0.62 D. The prevalence of With-the-rule, against the rule and oblique astigmatism was 93.8%, 4.7% and 1.5%, respectively. The mean anisometropia between two eyes was 0.14 ± 0.38 D. The most common type of refractive error was hyperopia (14.3%), followed by astigmatism (8.8%), anisometropia (3.2%), and myopia (0.9%). The refractive status in this population of urban Xuzhou preschool children was stable and there was no evidence of a myopic refractive shift over this age range in our cross-sectional study.     

Acute cytomegalovirus infection and the host immune response. I. Development and maintenance of cytomegalovirus (CMV) induced in vitro lymphocyte reactivity and its relationship to the production of CMV antibodies

Cytomegalovirus-induced lymphocyte reactivity (CMV-LR) of peripheral blood lymphocytes and antibodies against CMV-early (EA) and late (LA) antigens were studied in eighteen patients with documented acute CMV infections and seventy-six healthy persons. The development of a positive CMV-LR test lagged far behind the appearance of virus-specific antibodies. Positive CMV-LR tests were shown in all fourteen patients who could be tested twice or more. The median value of twenty-two tests in the acute phase (<50 days) was 269 d/min and the median of thirty tests in the post-illness phase (>50 days) was 1301 d/min (P<0·02). Once positive CMV-LR remained so during the follow-up period, up to 479 days after the onset of illness symptoms. In the meantime the LA and EA antibody response remained positive. Only two patients studied once showed negative CMV-LR responses while their serum contained high CMV-EA antibody titres. In CMV-LA seropositive healthy individuals who also possessed CMV-EA antibodies (LA+EA+) CMV-LR was higher (P<0·01) than in the LA seropositives who lacked EA antibodies (LA+EA−). The young LA+EA+ individuals, however, showed better (P<0·02) CMV−LR test results than the aged ones while their CMV antibody levels—especially the EA antibodies (P<0·02)—were lower. This phenomenon of increased CMV antibody production in relation to depressed CMV-LR is possibly caused by age-associated impairment of T lymphocyte function. The combined CMV-LR and EA antibody tests may provide useful means to study the specific immunological host/virus relationship in different clinical situations.     

Transport of potassium at the blood—brain barrier

1. The pial surface of different regions of the central nervous system of the rabbit have been bathed with artificial cerebrospinal fluid (c.s.f.), containing different concentrations of potassium. The object has been to change the composition of the interstitial fluid with respect to this ion, where it is adjacent to subarachnoid c.s.f.

2. Two techniques, subarachnoid perfusion from the supracallosal space between the hemispheres to the cisterna magna and barbotage from the cisterna magna, have been used. If the artificial c.s.f. contains Evans Blue, the former procedure results in maximum staining of the pia and underlying nervous tissue of the pons-medulla and spinal cord. The latter procedure results in maximum staining of the medial and supero-lateral surfaces of the cerebral hemispheres, particularly anteriorly.

3. During subarachnoid perfusion at 0·06 ml./min with the potassium-free fluid, most regions of the brain took up significantly greater amounts of ⁴²K than was the case during perfusion with the fluids containing 3 and 10 m-equiv/l. For blue cerebral cortex, the tissue subjected directly to the inflowing fluid and showing the biggest differences, the ratios, c.p.m. per g brain/c.p.m. per ml. plasma, were 1·71 ± 0·12 (+44%), 1·19 ± 0·05 and 1·07 ± 0·08 (-10%) during perfusion with the fluids containing, 0, 3 and 10 m-equiv/l. respectively.

4. During barbotage, the uptake of ⁴²K into pons-medulla and spinal cord from blood plasma, the concentration in the latter being effectively kept near constant, was, at the end of 2 hr, greater when the fluid contained potassium, 0 m=-equiv/l. rather than 10 m-equiv/l. Thus the ratio, c.p.m per g brain/c.p.m. per ml. plasma was (0·99 ± 0·04 (+36%) as against 0·73 ± 0·05 for pons-medulla where the difference was greatest.

5. Simultaneous measurements of the entry of [¹⁴C]urea from blood to different regions of the central nervous system revealed no significant differences due to the differing concentrations of potassium imposed by either barbotage or subarachnoid perfusion. This appears to exclude a non-specific cause for the changes in ⁴²K uptake, an example of which might be a changing blood flow.

6. Reasons are given for supposing that the big increase in ⁴²K uptake due to the potassium-free fluid must be due to events occurring at the blood—brain barrier. This might be some form of interaction, possibly the single file effect, such that a low potassium concentration in the interstitial fluid potentiates ⁴²K influx across the blood—brain barrier. Alternatively it might be due to a low potassium concentration in this fluid greatly reducing active potassium movement from interstitial fluid to blood. The former explanation would conform neatly with the present results; but the latter would additionally be compatible with other evidence concerning the homoeostasis of potassium concentration in c.s.f. and the interstitial fluid of brain.

     

Tumor cell and immune cell profiles in primary human glioblastoma: Impact on patient outcome

The distribution and role of tumor‐infiltrating leucocytes in glioblastoma (GBM) remain largely unknown. Here, we investigated the cellular composition of 55 primary (adult) GBM samples by flow cytometry and correlated the tumor immune profile with patient features at diagnosis and outcome. GBM single‐cell suspensions were stained at diagnosis (n = 44) and recurrence following radiotherapy and chemotherapy (n = 11) with a panel of 8‐color monoclonal antibody combinations for the identification and enumeration of (GFAP⁺CD45⁻) tumor and normal astrocytic cells, infiltrating myeloid cells —i.e. microglial and blood‐derived tumor‐associated macrophages (TAM), M1‐like, and M2‐like TAM, neutrophils. and myeloid‐derived suppressor cells (MDSC)— and tumor‐infiltrating lymphocytes (TIL) —i.e. CD3⁺T‐cells and their TCD4⁺, TCD8⁺, TCD4⁻CD8⁻, and (CD25⁺CD127^lo) regulatory (T‐regs) subsets, (CD19⁺CD20⁺) B‐cells, and (CD16⁺) NK‐cells—. Overall, GBM samples consisted of a major population (mean ± 1SD) of tumor and normal astrocytic cells (73% ± 16%) together with a significant but variable fraction of immune cells (24% ± 18%). Within myeloid cells, TAM predominated (13% ± 12%) including both microglial cells (10% ± 11%) and blood‐derived macrophages (3% ± 5%), in addition to a smaller proportion of neutrophils (5% ± 9%) and MDSC (4% ± 8%). Lymphocytes were less represented and mostly included TCD4⁺ (0.5% ± 0.7%) and TCD8⁺ cells (0.6% ± 0.7%), together with lower numbers of TCD4⁻CD8⁻ T‐cells (0.2% ± 0.4%), T‐regs (0.1% ± 0.2%), B‐lymphocytes (0.1% ± 0.2%) and NK‐cells (0.05% ± 0.05%). Overall, three distinct immune profiles were identified: cases with a minor fraction of leucocytes, tumors with a predominance of TAM and neutrophils, and cases with mixed infiltration by TAM, neutrophils, and T‐lymphocytes. Untreated GBM patients with mixed myeloid and lymphoid immune infiltrates showed a significantly shorter patient overall survival versus the other two groups, in the absence of gains of the EGFR gene (p = 0.02). Here we show that immune cell infiltrates are systematically present in GBM, with highly variable levels and immune profiles. Patients with mixed myeloid and T‐lymphoid infiltrates showed a worse outcome.     

Biophysical and mechanical properties of red and white muscle fibres in fish

1. The electrical properties of red muscle fibres of the silver carp (Carassius auratus (Linné)) were investigated and compared with those of white fibres. The resting membrane potential of red muscle was -73·1 mV, and of white muscle -82·4 mV. The effective resistance of fibre, time constant and space constant were 1·43 MΩ, 26·6 msec and 2·1 mm for red muscle and 1·0 MΩ, 48·4 msec and 1·9 mm for white muscle.

2. The membrane capacitance in red muscle was 2·55 μF/cm² and in white muscle was 7·23 μF/cm², though the sarcoplasmic reticulum and the transverse tubular system were developed equally in both tissues.

3. No rectification could be observed in the current—voltage relation in both the muscles. However, in excess potassium solutions, both types of muscle showed anomalous rectification.

4. In red muscle it was rare to trigger a spike by nerve stimulation or by field stimulation of the tissue. However, in white muscle, there appeared graded responses and occasionally spikes were generated with overshoot potentials.

5. Excess potassium concentration produced contracture in the red muscle, but even forty times of normal potassium never produced contracture in the white muscle.