Search

Essentials The underlying pathophysiological mechanisms behind cancer-associated thrombosis are unknown. We compared expression profiles in tumor cells from patients with and without thrombosis. Tumors from patients with thrombosis showed significant differential gene expression profiles. Patients with thrombosis had a proinflammatory status and increased fibrin levels in the tumor. Summary Background Venous thromboembolism (VTE) is a frequent complication in patients with cancer, and is associated with significant morbidity and mortality. However, the mechanisms behind cancer-associated thrombosis are still incompletely understood. Objectives To identify novel genes that are associated with VTE in patients with colorectal cancer (CRC). Methods Twelve CRC patients with VTE were age-matched and sex-matched to 12 CRC patients without VTE. Tumor cells were isolated from surgical samples with laser capture microdissection approaches, and mRNA profiles were measured with next-generation RNA sequencing. Results This approach led to the identification of new genes and pathways that might contribute to VTE in CRC patients. Application of ingenuity pathway analysis indicated significant links with inflammation, the methionine degradation pathway, and increased platelet function, which are all key processes in thrombus formation. Tumor samples of patients with VTE had a proinflammatory status and contained higher levels of fibrin and fibrin degradation products than samples of those without VTE. Conclusion This case-control study provides a proof-of-principle that tumor gene expression can discriminate between cancer patients with low and high risks of VTE. These findings may help to further unravel the pathogenesis of cancer-related VTE. The identified genes could potentially be used as candidate biomarkers to select high-risk CRC patients for thromboprophylaxis.

We report a case of pigmented villonodular synovitis (PVNS) of the hip joint in a patient with very limited impairment despite impressive radiological features. PVNS activates polyclonal, reactive macrophages with CSF1-receptors. Treatment options are discussed, including new possible therapies with CSF1-receptor inhibitors.

The anatomical distribution and the content of dendrite bundles were studied in the spinal cord of adult rats. We injected retrogradely transported Cholera Toxin subunit B into muscles in order to label motoneurons and their dendrites. Twenty-one muscles in the trunk and in extremities were selected which are involved in a variety of motor tasks. Reconstructions were made from consecutive sections and the organization of dendrites was studied. In addition, the distributions of fiber types in the muscles were estimated on the basis of ATPase and succinic dehydrogenase-stained sections. Dendrite bundles are confined to pools of muscles which have postural tasks. Two different types can be distinguished. In the first type, longitudinal dendrite bundles extend over the length of the pool and also transversal bundles occur. Such pools innervate the axial musculature, and the soleus and flexor carpi ulnaris muscles in the extremities. These muscles contain relatively high percentages of slow twitch type I muscle fibers. In the second type, the occurrence of bundles is confined to a few zones within the pool. Such pools innervate muscles which are involved in postural tasks but in addition they also subserve other movement tasks. Distinct regions within these muscles are characterized by high percentages of type I muscle fibers.

The EMG of the soleus muscle was recorded with bipolar electrodes chronically implanted in rats aged 10-30 days. Changes in the activity pattern were studied in relation to motor development. The firing pattern and shape of motor units were studied at higher resolution. EMG activity was closely related to motor behaviour and posture. At P11, soleus was only phasically active during movements. Tonic EMG activity appeared with age, and by P16 the activity pattern was similar to that of the adult. The activity level also increased markedly with age. This was paralleled by a change in hindlimb posture. The development of tonic activity precedes the development of dendrite bundles in the motoneuron pool of the soleus muscle, which does not support the hypothesis that bundle formation is causally related to tonic activity in postural muscles. At young ages, the tonic EMG was characterized by a clear oscillation pattern with a frequency ranging from 7 to 20 Hz, the higher frequencies occurring at older ages. This oscillation disappeared around P16, which suggests that it is related to functional development of the stretch reflex. Periods of high frequency firing were observed in the EMG; the highest firing frequencies occurred during locomotion. The maximum firing frequency increased steeply to about 80 Hz at P18. It is suggested that bursts of high frequency firing are related to the development of monoaminergic innervation of the spinal cord. During the first three weeks, motor unit action potentials often showed complex shapes of long duration and considerable spike-to-spike variability. Computer simulation of the summation process of fibre potentials at the electrode showed that the small fibre diameters and high variability in diameter at young ages are major factors with respect to the generation of complex potentials.

The development of the electromyogram (EMG) of tibialis anterior (TA) and medial gastrocnemius (GM) during locomotion was studied in normal rats from the onset of quadruped walking (postnatal day 10, P10) until P42. The objectives were to relate signal properties of the EMG and coordination of muscle activity to functional development of the hindlimb, which proceeds rapidly around P15. Both the EMG characteristics and the activation pattern showed marked changes with age. Initially, the EMG bursts were irregular and protracted. The activity level in the two muscles, in particular in GM, seemed to be low. Until P14, the motor units of GM showed a tendency towards synchronization. The EMG of TA consisted of an adult-like interference pattern from the youngest age studied. Although co-contraction of TA and GM was sometimes observed until P14, reciprocal activation of the muscles was evident at all ages. The timing of the alternating pattern became more accurate with age. The activity level in both muscles increased markedly from P15. These changes were reflected in the form factor (coefficient of variability) of the EMG and, to a lesser extent, in the power spectra. The time course of these changes bears a close temporal relationship to the development of locomotion. These results suggest that the degree of muscle activation is a decisive factor with respect to locomotor development. It is proposed that this is related to the maturation of supraspinal control.

The effects of undernutrition on the development of locomotion were studied in fourteen rats. Mothers received about 40% of normal quantities of standard laboratory food from the 5th day of gestation until weaning at the 21st day after birth. Qualitative as well as quantitative aspects of locomotion were studied from the 12th to the 30th day and compared to data from ten control rats. Undernutrition leads to delayed and prolonged developmental changes in locomotion. The adult type of locomotion, which in normal rats develops from the 14th day occurs after the 15th or 16th day in undernourished rats and this development also lasts longer. Locomotion remains mildly abnormal, at least until the 30th day. Especially at lower speeds, walking is clumsy because of an unusual brisk onset, a slightly unsteady gait and non-fluent paw movements. These results are discussed in the perspective of data on brain and muscle development in rats after early undernutrition.

The development of free walking was studied in rats between postnatal days 10 and 20. Spontaneous quadruped walking with the ventral surface of the body off the floor was first observed at postnatal day 11. Locomotion remained clumsy and invariably slow during the next few days, but a rapid transformation into the mature pattern of locomotion occurred around postnatal day 15. This transformation involved changes in quantitative parameters of locomotion as well as a change in the movement pattern of the hindlimb. A swimming-like movement characterized by abduction, rotation and hyperextension of the paw was replaced by the digitigrade adult pattern without marked rotation. Joint angle trajectories of the major joints during the step cycle changed considerably during the transitional period. The results, which are to serve as a framework for ongoing research into the effects of early undernutrition and movement restriction upon motor function, are discussed in the perspective of developmental changes in the nervous and musculoskeletal system.

Storage of oligosaccharides due to a deficiency of alpha-mannosidase can lead to joint destruction in children and young adults. Treating hip destruction with a prosthesis might be successful in some of these patients, although diminished bone quality increases the risk of loosening of the prosthesis.

The degree of healing and damage of the bronchial wall after photodynamic therapy, Nd-YAG laser and electrocautery for intraluminal early-stage cancer have been analysed. Review of the bronchoscopy reports and follow-up histology specimens of twenty-nine patients treated bronchoscopically with curative intent for their intraluminal tumor have been performed. Seventeen patients had been treated with bronchoscopic electrocautery (BE) only, six with photodynamic therapy (PDT) and six with Nd-YAG laser. Bronchial wall scarring seen during follow-up bronchoscopy was scored and subepithelial fibrosis were histologically evaluated using Alcian blue staining, Azan staining and polarised light. After BE, prominent airway scarring was seen in five patients (29%), with significant stenosis (>50% lumen) in one of these cases. Prominent scarring and significant stenosis were found in four (67%), after PDT. In five (83%) after Nd-YAG laser prominent scarring was found, one patient had significant stenosis. In three cases, two after BE and one after PDT, subepithelial tissue in the follow-up biopsies was insufficient for proper histologic examination. In the remaining biopsy specimen only one (7%) showed a moderate or excessive amount of fibroblasts after BE, whereas for PDT and Nd-YAG this was found in three (60%) and four patients (67%), respectively. Excessive matrix was found in none of the biopsies after BE, in two (40%) after PDT and in three (50%) after Nd-YAG laser. Compact collagen formations were seen in two (12%) biopsies after BE, in two (40 and 33%) after PDT and Nd-YAG, respectively. Compared to electrocautery, more airway scarring and more subepithelial fibrosis were seen after treatment with PDT and Nd-YAG laser. These findings, especially regarding PDT, is in contrast to the assumption that PDT is selective and may be important in the choice of treatment for patients with early stage cancer.

* Professor J M M Walboomers died recently Background/Aims—To improve the accuracy of conventional cytology in cervical cancer screening, high risk human papillomavirus (HPV) testing and neural network based screening have been developed. This study assessed the power of both techniques to detect women at risk of developing incident CIN III; that is, CIN III detected during the follow up of women with normal cytology and borderline nuclear changes. Methods—A cohort of 2250 women, 34–54 years of age, who attended population based cervical cancer screening from 1988 to 1991 and had normal smears or borderline nuclear changes was followed. All smears were tested for high risk HPV and the smears were rescreened using neural network based screening. The value of neural network based screening for predicting incident CIN III during a mean follow up period of 6.4 years was compared with that of high risk HPV testing. In addition, morphological markers presumed to be related to HPV were correlated with HPV status. Results—Thirteen (0.6%) women had incident CIN III. Both high risk HPV positivity and abnormal cytology were associated with an increased risk for incident CIN III (odds ratio, 240 and 22, respectively) and high risk HPV positivity was associated with abnormal cytology. The sensitivity of high risk HPV testing for predicting incident CIN III was much higher than that of neural network based screening (92% and 46%, respectively). None of the morphological markers assessed, including koilocytosis, was correlated with high risk HPV status. Conclusion—High risk HPV testing is superior to neural network based screening in identifying women at risk of developing CIN III. For women with normal cytology and borderline changes and a negative high risk HPV test, the screening interval can be considerably prolonged. Key Words: neural network based screening • high risk human papillomavirus testing • CIN III

Patients with advanced stages of head and neck cancer frequently develop locoregional recurrence as well as distant metastases. These data indicate that traditional diagnostic methods such as histopathology and radiology are not sensitive enough to detect the small numbers of tumor cells which are left behind, defined as minimal residual disease (MRD). Sensitive diagnostic assays based on molecular markers appear to be powerful tools to improve the staging of these patients. At the DNA level, tumor-specific p53 mutations seem to have great potential for the detection of "occult" tumor cells at surgical margins and lymph nodes. At the RNA level HNSCC associated antigens like the E48 antigen, allow the detection of rare HNSCC cells in blood and bone marrow and, it is hoped, also in lymph nodes and lymph node aspirates. However, the molecular assays which are used to detect MRD are subject to certain (technical) problems which affect their sensitivity and specificity. In this paper we will present examples of molecular assays such as the plaque assay using p53 mutations and the E48 RT-PCR, and show their use for MRD detection in cervical lymph nodes. In addition, we will discuss the problems and pitfalls associated with these sensitive techniques.

To study tissue effects of bronchoscopic electrocautery (BE).In six patients with non-small cell lung cancer, a BE procedure was performed immediately before surgery. After patients were placed on ventilation, normal mucosa on different carinae was treated with a cautery probe (2-mm(2) surface area) at a power setting of 30 W with a variable time of application of 1 to 5 s. Bronchoscopic appearance of the treated area was documented photographically, and histologic changes of the bronchial wall were examined.Bronchoscopy unit of a university hospital.BE resulted bronchoscopically in whitening of the bronchial mucosa with crater-shaped lesions. After longer duration of BE application, deeper craters with more profound charring were seen. Histologic changes of the lesions showed craters containing a variable amount of necrotic tissue. In one case, thin subsegmental carinae were coagulated and measurements could not be performed. In the remaining five cases, microscopic findings revealed 0.2 +/- 0.1-mm necrosis after 1 s; 0.4 +/- 0.2-mm necrosis after 2 s; 0.9 +/- 0.5-mm necrosis after 3 s; and 1.9 +/- 0.8-mm necrosis after 5 s. A variable degree of tissue damage surrounding the necrotic tissue area was found. In one case, cartilage damage appeared after 3 s of coagulation, and extensive damage of the underlying cartilage was seen in four cases after 5 s of application.Superficial damage was obtained by short duration of BE (or = 2 s), and longer duration of coagulation (3 s or 5 s) caused damage to the underlying cartilage. Bronchoscopic appearance after endobronchial electrocautery corresponded with the histologic changes.

Motoneuronal pools of muscles that subserve postural tasks contain dendrite bundles. We investigated in the rat the development of these bundles in the pools of the long back muscles and related this to postural development. Motoneurons and their dendrites were retrogradely labeled by injecting unconjugated cholera toxin subunit B (CTB) into the muscles of 54 normal rats from birth until adulthood and into 18 rats that were vestibularly deprived from the 5th postnatal day (P5). Dendrite bundles coursing in a transverse direction already occurred at P1. From P4, the first longitudinal bundles could be observed, but the major spurt in development occurred between P6 and P9, when conspicuous bundles developed coursing in rostro-caudal and tranverse directions. This is the age when rats become able to stand freely and walk a few steps. Around P20, the dendrite bundles attained their adult characteristics. Vestibular deprivation by plugging both semicircular horizontal canals did not lead to a retarded development of dendrite bundles nor to a changed morphology. This finding is remarkable, as behavioral analysis showed a delay in postural development by about 3 days. We hypothesize that dendrite bundles in the pools of the long back muscles function to synchronize the motoneurons in different spinal cord segments.

Patients with advanced stages of head and neck cancer frequently develop locoregional recurrence as well as distant metastases. These data indicate that traditional diagnostic methods such as histopathology and radiology are not sensitive enough to detect the small numbers of tumor cells which are left behind, defined as minimal residual disease (MRD). Sensitive diagnostic assays based on molecular markers appear to be powerful tools to improve the staging of these patients. At the DNA level, tumor-specific p53 mutations seem to have great potential for the detection of "occult" tumor cells at surgical margins and lymph nodes. At the RNA level HNSCC associated antigens like the E48 antigen, allow the detection of rare HNSCC cells in blood and bone marrow and, it is hoped, also in lymph nodes and lymph node aspirates. However, the molecular assays which are used to detect MRD are subject to certain (technical) problems which affect their sensitivity and specificity. In this paper we will present examples of molecular assays such as the plaque assay using p53 mutations and the E48 RT-PCR, and show their use for MRD detection in cervical lymph nodes. In addition, we will discuss the problems and pitfalls associated with these sensitive techniques.

The development of the EMG of the multifidus (MM) and longissimus lateralis (LL) muscles was studied in 16 rats between the 6th day (P6) and P45 and related to behavioural development. The EMG changes gradually from irregular and spiky towards a regular interference pattern and simultaneously, the general activity level increases. From P11, tonic background activity occurs with phasic activity superimposed. During locomotion, a relation between increasing activity in the long back muscles and the swing phase of both hind paws develops, but only from P15 the activity modulates consistently with the step cycle. From the end of the second week of life, the activity in the long back muscles precedes hind paw movements. The development of the EMG of the LL and MM is closely followed by the development of adult and fluent motor patterns. This suggests that the stabilisation of the trunk is essential for the development of these patterns.

The effect of early immobilization upon the adult locomotor pattern was studied. One hindlimb of neonatal rats was immobilized during 20 days and the EMG pattern was studied 3-8 weeks after termination of movement restriction. All rats showed a fluent locomotion pattern at these ages, but the EMG pattern revealed differences in the timing of flexor and extensor activity between the previously immobilized and the contralateral side. We suggest that interference with movement related afferent feedback at young ages has subtle but long-lasting effects upon the locomotion generating neural circuits.

The development of the nervous system is determined by an interaction between genetic and epigenetic factors. We investigated the possible role of proprioceptive afferent input in the development of locomotion in the rat. Kinematic analysis of locomotion in normal rats reveals a marked transition from immature overground locomotion into the adult pattern around the 15th postnatal day. Around this age, the timing of EMG activity in the tibialis anterior and the gastrocnemius muscle in the hindpaw gains accuracy and the intensity of the EMG increases. In the soleus muscle we observbed an increased regularity in the shape of individual motor unit potentials. Neuroanatomical research revealed the occurrence of dendrite bundles in the motoneuronal pool of the soleus muscle pool from the ]6th day as the result of a reorganisation. Immobilisation of one hindlimb in an extended position by casts around the leg from the first till the 20th postnatal day does not interfere with the development of dendrite bundles in the soleus motoneuronal pool. Howeverm, we demonstrated long lasting abnormalities in the timing of the EMG activation patterns in the gastrocnemius and the tibialis anterior muscles. It is conceivable that the experimental interference with afferent feed back is the causative factor of these functional abnormalities.

Postural development has not attracted much attention in investigations of neuro-ontogeny. In the rat, motor behaviours have been studied repeatedly but the development of postural control has been largely neglected. In the present study we have taken inventory of behavioural aspects of postural development. Postures and postural skills of 10 pups were 3-dimensionally recorded on video tape from the 2nd to the 20th day. The development of posture may be subdivided into three periods. During the first period, lasting until the 4th or 5th day, neonates were unable to lift their trunk from the floor, and the only indication for postural activity being head lifting. From then until the 12th-13th day, pups were able to walk, though staggering, and in addition they groomed and reared with forelimb support. From then, in a rapid development lasting until day 16, the adult type of fluent locomotion developed as well as grooming and rearing without support. In addition, complex motor acts developed. These results are discussed in the perspective of the structural and functional development of postural systems.

4036 Background: Between 25 and 35% of stage II CRC patients will experience a recurrence of their disease and may benefit from adjuvant chemotherapy. Official guidelines give suggestions but no clear recommendation for best risk stratification. Here we describe the development a robust signature that predicts disease relapse and can assist in treatment decisions. Methods: Fresh frozen tumor tissues from 180 patients with stage I, II and III colorectal cancer undergoing surgery were analyzed using high density Agilent 44K oligonucleotide arrays. Median FU was 70.2 months; 85% of patients did not receive adjuvant chemotherapy. Unsupervised hierarchical clustering based on full-genome gene expression measurement indicated the existence of 3 main colon molecular subclasses. Survival analysis of the 3 classes showed that subtype C (n= 27) had a poor outcome and subtype A (n= 48) good outcome. Only the intermediate group B (n=104) was used to develop a signature by using a cross validation procedure to score all genes for their association with 5-yr distant metastasis free survival (DMFS) and subsequently applied to all samples (n=180). The obtained gene signature was further validated on an independent cohort of 178 stage II + III colon samples. Results: A set of 38 prognosis related gene probes showed robust DMFS association in over 50% of all iterations in the Training Set of 180 samples. The gene signature was validated on an independent cohort of 178 samples from stage II + III colon cancer patients. The profile classified 61% of the validation samples as low-risk and 39% as high-risk. The low- and high-risk samples showed a significant difference in DMFS with a HR of 3.19 (P= 8.5e-4). Five-year DMFS rates were 89% (95%CI 83–95) for low-risk and 62% (95%CI 50–77) for high-risk samples. Moreover, the profile showed a significant performance within stage II (P=0.0058) and III (P=0.036) only samples. The performance of the profile was significant for both untreated (P=0.0082) and treated patients (P=0.016) suggesting that its power is independent of treatment benefits. Conclusions: ColoPrint is able to predict the prognosis of stage II and III colon cancer patients and facilitates the identification of patients who would benefit from adjuvant chemotherapy. [Table: see text]

A panel of monoclonal antibodies directed against various lymphoid and non-lymphoid cell subsets was used to study the lymph nodes of human fetuses of 16-40 weeks. B cells were of intermediate size and were present at all ages in primitive follicles and in the outer cortex. The fetal B-cell immunophenotype is indicative of an intermediate stage of development, just preceding the differentiation to mature B cell. Forty to sixty per cent Leu1+ B cells were observed in the follicles until the end of the second trimester. At all stages, T cells showed an immunophenotype similar to type III thymocytes, different from adult peripheral T cells, with a marked predominance of CD4+ T cells. Leu7+ NK cells were generally absent. OKIa+ interdigitating reticulum cells were present in T-cell areas. Some axillary lymph nodes showed strongly CD1+ dendritic cells, probably Langerhans' cells. Macrophages and granulocytes were present in varying numbers. Altogether, our results indicate that fetal lymph nodes are quite well differentiated at an early fetal age, although T and B cells do not (yet) show adult immunophenotypes. The expression of the CD38 antigen may be a main marker related to the immaturity of fetal T and B cells.