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Most patients with triple-negative breast cancer (TNBC) are not candidates for targeted therapy, leaving chemotherapy as the primary treatment option. Recently, immunotherapy has demonstrated promising results in TNBC, due to its immunogenicity. In addition, a novel antibody-drug conjugate, namely, trastuzumab-deruxtecan, has shown effectiveness in TNBC patients with low-HER2 expression (HER2-low). These novel treatment options raise the question about the potential association between the density of stromal tumor-infiltrating lymphocytes (sTILs) and the level of HER2 expression. We aimed to evaluate the association between the level of HER2 expression (HER2-low versus HER2-0) and density of sTILs in TNBC patients, and how they impact the response to neoadjuvant chemotherapy (NAC). This was a retrospective multicenter study including all TNBC patients diagnosed between 2018 and 2022. Central pathology review included sTILs percentages and level of HER2 expression. Tumors were reclassified as either HER2-0 (HER2 IHC 0) or HER2-low (IHC 1 + or 2 + with negative reflex test). Various clinicopathologic characteristics, including sTILs density, and response to NAC were compared between HER2-0 and HER2-low cases. In total, 753 TNBC patients were included in this study, of which 292 patients received NAC. Interobserver agreement between the original pathology report and central review was moderate (77% had the same IHC status after reclassification in either HER2-0 or HER2-low; k = 0.45). HER2-low TNBC represented about one third (36%) of the tumors. No significant difference in sTILs density or complete pathologic response rate was found between HER2-0 and HER2-low cases (p = 0.476 and p = 0.339, respectively). The density of sTILs (≥ 10% sTILs vs. < 10%) was independently associated with achieving a pCR (p = 0.011). In conclusion, no significant association was found between HER2-low status and density of sTILs nor response to NAC. Nonetheless, sTILs could be an independent biomarker for predicting NAC response in TNBC patients., (© 2024. The Author(s).)

Tumor-associated macrophages (TAMs) and microglia (MG) are potent regulators of glioma development and progression. However, the dynamic alterations of distinct TAM populations during the course of therapeutic intervention, response, and recurrence have not yet been fully explored. Here, we investigated how radiotherapy changes the relative abundance and phenotypes of brain-resident MG and peripherally recruited monocyte-derived macrophages (MDMs) in glioblastoma. We identified radiation-specific, stage-dependent MG and MDM gene expression signatures in murine gliomas and confirmed altered expression of several genes and proteins in recurrent human glioblastoma. We found that targeting these TAM populations using a colony-stimulating factor-1 receptor (CSF-1R) inhibitor combined with radiotherapy substantially enhanced survival in preclinical models. Our findings reveal the dynamics and plasticity of distinct macrophage populations in the irradiated tumor microenvironment, which has translational relevance for enhancing the efficacy of standard-of-care treatment in gliomas., (Copyright © 2020 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.)

Background: Selective removal of initially tumor-positive axillary lymph nodes in breast cancer patients who underwent neoadjuvant systemic treatment (NST) improves the accuracy of nodal staging and provides the opportunity for more tailored axillary treatment. This study evaluated whether radioguided occult lesion localization (ROLL) of clip-marked lymph nodes is feasible in clinical practice., Methods: Prior to NST, a clip marker was placed inside a proven tumor-positive lymph node in all breast cancer patients (cTis-4N1-3 M0). After NST, technetium-99m-labeled macroaggregated albumin was injected in the clip-marked lymph nodes. The next day, these ROLL-marked nodes were selectively removed at surgery to evaluate the pathological response of the axilla., Results: Thirty-seven patients (38 axillae) underwent clip insertion. After NST, the clip was visible by ultrasound in 36 procedures (95%). In the other two patients, the ROLL-node injection was performed in a sonographically suspicious unclipped node (1), and near the clip under computed tomography guidance (1). Initial surgery successfully identified the ROLL-marked node with clip in 33 procedures (87%). Removed specimens in the other five procedures contained only the sonographically suspicious tumor-positive unclipped node (1), a node with signs of complete response but no clip (2), a clip without node (1), and tissue without node nor clip, and a second successful ROLL-node procedure was performed (1). Overall, 10 ROLL-marked nodes had no residual disease., Conclusions: This study demonstrates that the ROLL procedure to identify clip-marked lymph nodes is feasible. This facilitates selective removal at surgery and may tailor axillary treatment in patients treated with NST.

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., (© 2017 International Society on Thrombosis and Haemostasis.)

Background: Moderate diagnostic accuracy of MRI and initial cerebrospinal fluid (CSF) cytology analysis results in at least 10%-15% false negative diagnoses of leptomeningeal metastases (LM) of solid tumors, thus postponing start of therapy. The aim of this prospective clinical study was to determine the diagnostic value of epithelial cell adhesion molecule (EpCAM)-based flow cytometry versus cytology in CSF for the diagnosis of LM in patients with epithelial tumors., Methods: Patients with a clinical suspicion of LM but a negative or inconclusive MRI in whom a diagnostic lumbar puncture has to be performed were included. At least 5 mL of CSF for cytology, 5 mL for flow cytometry, 2 mL for cell count and biochemistry, and 8 mL whole blood samples for circulating tumor cells measurements and biochemistry were drawn. Tumor cells in CSF and whole blood were detected by multiparameter flow cytometry using EpCAM antibody., Results: In total 29 eligible patients were enrolled in the study. Thirteen patients were ultimately diagnosed with LM. The flow cytometry assay showed 100% sensitivity and 100% specificity for diagnosing LM, while sensitivity of CSF cytology was only 61.5%. Cell count or biochemical parameters in CSF were abnormal in 100% of patients with LM., Conclusions: Our results suggest that the EpCAM-based flow cytometry assay is superior to CSF cytology for the diagnosis of LM in patients with an epithelial tumor, a clinical suspicion of LM, and a nonconclusive MRI. Confirmation of these data is needed in a larger dataset to recommend dual CSF diagnostics for LM., Clinicaltrialsgov Identifier: NCT01713699., (© The Author(s) 2015. Published by Oxford University Press on behalf of the Society for Neuro-Oncology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Background: Osteolytic cyst formation after total ankle arthroplasty has been identified in recent years and is probably an important problem with longer follow-up. The aim of this study is to describe the outcome of a histological analysis of samples from periprosthetic intra-osseous cysts and joint capsules, retrieved during revision surgery after primary total ankle arthroplasty., Methods: All samples (n=22) were analyzed and scored using a semi-quantitative grading system. The main items of interest were polyethylene (PE) particles, metal particles, histiocytes, and giant cells., Results: All cyst samples contained PE particles. A similar number of PE particles were found in talar and tibial cysts. No significant correlation between the number of PE particles and time to reoperation was found. Metal particles were found in 16 cysts., Conclusions: We suggest that PE particles are not the primary cause of osteolytic cyst formation but a secondary contributing factor probably accelerating the process of osteolysis. It is likely that implant design, biomechanical factors and local anatomic-physiological factors play an important role., (Copyright © 2015 European Foot and Ankle Society. Published by Elsevier Ltd. All rights reserved.)

Introduction: Sarcoidosis is a granulomatous, multisystem inflammatory disease of unknown cause, which presents with a wide variety of symptoms. We describe a rare case of a newly diagnosed sarcoidosis, with cluster-like headache as a presenting symptom., Case: A 31-year-old man presented with cluster headache with a cystic lesion in the hypothalamus. A non-caseating granuloma consistent with the diagnosis sarcoidosis was found at biopsy. Pulmonary involvement was confirmed on positron electron tomography-computed tomography (PET-CT). Treatment with prednisone led to regression of the hypothalamic lesion. Headache attacks did not recur., Discussion: Cluster-like headache with a cystic hypothalamic lesion as first presentation of sarcoidosis has never been reported. Their possible relationship seems to underline the role of the hypothalamus in the central pain-regulatory areas in the brain, but is not undisputed. This case clearly demonstrates once again the relevance of neuroimaging in new-onset cluster-like headache.

Purpose: This study aims to develop a robust gene expression classifier that can predict disease relapse in patients with early-stage colorectal cancer (CRC)., Patients and Methods: Fresh frozen tumor tissue from 188 patients with stage I to IV CRC undergoing surgery was analyzed using Agilent 44K oligonucleotide arrays. Median follow-up time was 65.1 months, and the majority of patients (83.6%) did not receive adjuvant chemotherapy. A nearest mean classifier was developed using a cross-validation procedure to score all genes for their association with 5-year distant metastasis-free survival., Results: An optimal set of 18 genes was identified and used to construct a prognostic classifier (ColoPrint). The signature was validated on an independent set of 206 samples from patients with stage I, II, and III CRC. The signature classified 60% of patients as low risk and 40% as high risk. Five-year relapse-free survival rates were 87.6% (95% CI, 81.5% to 93.7%) and 67.2% (95% CI, 55.4% to 79.0%) for low- and high-risk patients, respectively, with a hazard ratio (HR) of 2.5 (95% CI, 1.33 to 4.73; P = .005). In multivariate analysis, the signature remained one of the most significant prognostic factors, with an HR of 2.69 (95% CI, 1.41 to 5.14; P = .003). In patients with stage II CRC, the signature had an HR of 3.34 (P = .017) and was superior to American Society of Clinical Oncology criteria in assessing the risk of cancer recurrence without prescreening for microsatellite instability (MSI)., Conclusion: ColoPrint significantly improves the prognostic accuracy of pathologic factors and MSI in patients with stage II and III CRC and facilitates the identification of patients with stage II disease who may be safely managed without chemotherapy.

Aim: Novel diagnostic breast cancer markers have been extensively searched for in the proteome, using, among others, surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS). Thus far, the majority of SELDI-TOF MS studies have investigated samples originating from biorepositories, which hampers biomarker discovery as they likely suffer from variable adherence to collection protocols., Material and Methods: We investigated breast cancer (n=75) and control (n=26) serum and tissue samples, collected prospectively by rigorous adherence to a strictly defined protocol. Sera were collected preoperatively and postoperatively, and serum and tissue samples were analyzed by SELDI-TOF MS using the IMAC30 Ni and Q10 pH 8 array., Results: Three serum peaks were significantly associated with breast cancer, while in tissue, 27 discriminative peaks were detected. Several peak clusters gradually increased or decreased in intensity from healthy to benign to cancer, or with increasing cancer stage. The constructed classification trees had a tenfold cross-validated performance of 67% to 87%. Two tissue peaks were identified as N-terminal albumin fragments. These are likely to have been generated by (breast) cancer-specific proteolytic activity in the tumor microenvironment., Conclusions: These albumin fragment scan potentially provide insights into the pathophysiological mechanisms associated with, or underlying, breast cancer, and aid in improving breast cancer diagnosis.

Background: HIV-infected women are at increased risk for cervical dysplasia. Cervical dysplasia is caused by persistent infections with certain types of human papillomavirus (HPV). Conventional testing for genital HPV infections requires cervical cytology. A non-invasive screening method by detection of HPV DNA in urine samples is preferable but is not a routine practice., Objectives: To investigate the prevalence and concordance of HPV in paired urine and cervical smear samples and cytological results of Pap smears in HIV-infected women., Study Design: Paired urine and cervical smear samples were collected from 27 HIV-infected women., Results: The HPV prevalence in urine and cervical smear samples was 81.5% and 51.9%, respectively (p = 0.01). The concordance for HPV positivity and negativity between urine and cervical smear samples is 71%. Seven women (25.9%) had an abnormal cervical smear of Pap II or higher. In all urine samples from these cases HPV DNA was detected., Conclusion: In the present study we show that the HPV prevalence in urine and cervical smear samples of HIV-infected women is high and HPV test results are highly concordant. Therefore, urine samples can be used as medium for HPV testing. HPV testing in urine samples is a simple, reliable, non-invasive screening method.

Objective: Nodular regenerative hyperplasia (NRH) and sinusoidal dilatation have been described in relation to thiopurine use in patients with inflammatory bowel disease (IBD). However, there is a dearth of data on the prevalence of these histological abnormalities in general. The aim of our study was to describe the prevalence of these histological liver changes in a thiopurine-naïve IBD cohort., Material and Methods: Liver biopsy specimens were obtained from patients who were treated in a referral center and who underwent gastrointestinal surgery for IBD. Patients were excluded if thiopurines were ever used. The liver specimens were pathohistologically assessed with special attention to NRH., Results: A total of 83, properly stained, liver specimens (Crohn's disease 61%) were evaluated. NRH was observed in 6% compared to sinusoidal dilatation of varying degree in 34% of specimens. An older age at biopsy was correlated with NRH (p=0.015). Fibrosis and steatosis of varying degrees were detected in 31% and 36% of liver biopsies, respectively. No cases of liver cirrhosis were observed., Conclusions: Pathohistological hepatic abnormalities are common in non-thiopurine using IBD patients. The association between thiopurine use, NRH and sinusoidal dilatation may be weaker than as reported in recent literature, as there is relatively high background prevalence in selected series.

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.

Gastric carcinogenesis is driven by an accumulation of genetic changes that to a large extent occur at the chromosomal level. We analysed the patterns of chromosomal instability in 35 gastric carcinomas and their clinical correlations. With microarray competitive genomic hybridization, genomewide chromosomal copy number changes can be studied with high resolution and sensitivity. A genomewide scanning array with 2275 BAC and P1 clones spotted in triplicate was used. This array provided an average resolution of 1.4 Mb across the genome. Patterns of chromosomal aberrations were analysed by hierarchical cluster analysis of the normalized log(2) tumour to normal fluorescence ratios of all clones, and cluster membership was correlated to clinicopathological data including survival. Hierarchical cluster analysis revealed three groups with different genomic profiles that correlated significantly with lymph node status (P=0.02). Moreover, gastric cancer cases from cluster 3 showed a significantly better prognosis than those from clusters 1 and 2 (P=0.02). Genomic profiling of gastric adenocarcinomas based on microarray analysis of chromosomal copy number changes predicted lymph node status and survival. The possibility to discriminate between patients with a high risk of lymph node metastasis could clinically be helpful for selecting patients for extended lymph node resection.

Basal cell carcinoma of the anus is extremely rare, accounting for only 0.1% of anorectal malignancies. We report the reconstruction of the anus with a biaxial pedicled scrotal island myofasciocutaneous flap following excision of an anal basal cell carcinoma, and conclude that this technique may be valuable in the reconstruction of the anal region.

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.

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.

Study Objectives: To study tissue effects of bronchoscopic electrocautery (BE)., Design: 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., Setting: Bronchoscopy unit of a university hospital., Measurements and Results: 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., Conclusions: 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.

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.

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 transverse 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.

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 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 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 observed 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 16th 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. However, 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.

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.

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.

The morphological development of motoneuron pools of two hindlimb muscles of the rat, soleus (SOL) and tibialis anterior (TA), was studied in rats ranging in age between 8 and 30 postnatal days (P8-P30). Motoneurons were retrogradely labelled by injecting a cholera toxin B subunit solution directly into the muscles. This resulted in extensive labelling of motoneurons as well as their dendritic trees. The distribution of cross sectional areas of neuronal somata was determined for both muscles at various ages. Somal size increased considerably between P8 and P12, whereas growth was moderate between P12 and P20. The size distribution of SOL motoneurons was bimodal from P20, whereas the size distribution of TA motoneurons remained largely unimodal. The morphological development of the dendritic tree was studied qualitatively. The development of dendritic arborization within the SOL and the TA motoneuron pool showed major differences. The arborization pattern of dendrites of TA motoneurons was basically multipolar at all ages. In contrast, dendrites of SOL neurons tended to line up with the rostro-caudal axis and became organized in longitudinal bundles from P16 onwards. The relatively late appearance of dendrite bundles in the soleus motoneuron pool suggests that they might be related to the fine-tuning of neuronal activity rather than patterning of motor activity. The occurrence of dendrite bundles in SOL and not in TA motoneuron pools suggests that they may be related to the different afferent organization of this postural muscle or to its tonic activation pattern.

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.

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.