Breast cancer during follow-up and progression - A population based cohort on new cancers and changed biology.

BACKGROUND: Emerging data indicate an important role for biopsies of clinically/radiologically defined breast cancer 'recurrences'. The present study investigates tumour related events (relapses, other malignancies, benign conditions) after a primary breast cancer diagnosis. PATIENTS AND METHODS: The cohort includes 2102 women, representing all patients, with primary invasive breast cancer during 2000-2011 in the county of Värmland, Sweden. A comparative analysis of oestrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2 (HER2) and proliferation (Ki67) between the primary tumour and the relapse was performed and related to outcome. RESULTS: With a mean follow-up time of 4.8 years, 1060 out of 2102 patients have had a biopsy taken after the initial breast cancer diagnosis demonstrating 177 recurrences, 93 other malignancies (colorectal, lung, skin), 40 cancer in situ (skin, breast) and 857 benign lesions. Approximately 70% (177 out of 245) of all cases of relapsed breast cancer underwent a biopsy during this time period. For patients with recurrences, ER (n=127), PR (n=101), HER2 (n=73) and Ki67 (n=55) status in both primary tumour and the corresponding relapse were determined. The discordance of receptor status was 14.2%, 39.6%, 9.6% and 36.3%, respectively. Loss of ER or PR in the relapse resulted in a significant increased risk of death (hazard ratio (HR) 3.62; 95% confidence interval (CI), 1.65-7.94) and (HR 2.34; 95% CI, 1.01-5.47) compared with patients with stable ER or PR positive tumours. The proportion of patients losing ER was bigger in the group treated with endocrine therapy alone or in combination with chemotherapy, 16.7% and 13.3%, respectively, compared with the group treated with chemotherapy alone or that which received no treatment 4.3% and 7.7%, respectively. CONCLUSION: Discordance of biomarkers between the primary tumour and the corresponding relapse was seen in 10-40% of the patients, adjuvant therapies seem to drive clonal selections. Patients with tumours losing ER or PR during progression have worse survival compared with patients with retained receptor expression.

Spontaneous bursting neuronal discharges recorded from peripheral nerve in human: injury discharges or not?

This paper deals with a spontaneous, bursting neuronal activity which can not be altered by any stimulation in the periphery or voluntary actions or by cognitive tasks. An initial description of such units led to the conclusion that this activity was generated ectopically at the site of a previous or present impalement of a nerve fibre. The aim of the current study was to record a larger number of these units by using microneurography, in order to characterise their firing properties and particularly, see if any subtypes of units could be identified. In conclusion, this paper suggests that some of these discharges could be related to an injury of the nerve fibre, however most of them could not. Some hypothesis regarding the nature of these bursting activities are suggested.

Influences on the fusimotor-muscle spindle system from chemosensitive nerve endings in cervical facet joints in the cat: possible implications for whiplash induced disorders.

The aim of the present study was to establish if there exists reflex connections from ligamentous structures in cervical facet joints and the fusimotor system of dorsal neck muscles. In seven cats, anaesthetized with alpha-chloralose, bradykinin (BK) of concentrations between 12 and 50 microg was injected into the facet joint between C1 and C2. Recordings were made from single muscle spindle afferents (MSA) originating in contralateral trapezius and splenius muscles (TrSp). Fusimotor induced changes in the sensitivity of the muscle spindle afferents were assessed by recording the responses to sinusoidal stretches of the TrSp muscles. The mean rate of discharge and the depth of modulation of a fitted sine were taken as quantitative estimates of the response. A total of 25 MSAs were recorded, and 21 of these showed clear-cut alterations in their responses to the sinusoidal stretches following Bk. injections into contralateral facet joint. The majority of the responding afferents (13/21) showed changes in their responses indicating an increased activity of static fusimotoneurones, although responses of dynamic and mixed static and dynamic nature were also seen. Local anaesthetics applied to the intraarticular receptors abolished the effects. Injection (i.v.) of a general anaesthetic (pentobarbital) abolished the effects. The results show that there exist reflex connections between receptors in cervical facet joints and fusimotoneurones of dorsal neck muscles, and this might be of importance in the pathophysiology behind whiplash associated disorders (WAD).

Elevated intramuscular concentration of bradykinin in jaw muscle increases the fusimotor drive to neck muscles in the cat.

Patients suffering from temporomandibular dysfunction exhibit clinical findings such as fatigue, painful muscles, and muscles that are tender to palpation, not only in the temporomandibular area, but also in the neck/shoulder region. The mechanisms behind this are not known, although previous studies of similar spreading phenomena have revealed a possible involvement of the fusimotor system. In the present study, we evaluated the activity of this system by recording the activity of muscle spindle afferents from dorsal neck muscles after intramuscular injections of 6 microg to 25 microg bradykinin in the ipsilateral masseter muscle. A total of 23 muscle spindle afferents from the trapezius and splenius muscles was recorded at the C3-C4 level in 7 adult cats anesthetized with alpha-chloralose. Of these 23 afferents, 17 (74%) showed significantly different responses to bradykinin injections compared with control injections (dissolving agent, Tyrode), and the majority of the effects were compatible with an increased static fusimotor drive to the muscle spindle system. Thus, the results demonstrate potent reflex connections from groups III and IV masseter muscle afferents to fusimotor neurones on the C3-C4 level. It is concluded that the fusimotor system might play a significant role in the mechanisms behind the spread of muscle pain and tension from the temporomandibular region to the neck.

Proprioceptive population coding of two-dimensional limb movements in humans: I. Muscle spindle feedback during spatially oriented movements.

The proprioceptive coding of multidirectional ankle joint movements was investigated, focusing in particular on the question as to how accurately the direction of a movement is encoded when all the proprioceptive information from all the muscles involved in the actual movement is taken into account. During ankle movements imposed on human subjects, the activity of 30 muscle spindle afferents originating in the extensor digitorum longus, tibialis anterior, extensor hallucis longus and peroneus lateralis muscles was recorded from the lateral peroneal nerve using the microneurographic technique. In the first part of the study, it was proposed to investigate whether muscle spindle afferents have a preferred direction, as previously found to occur in the case of cortical cells, and to analyze the neural coding of the movement trajectories using a "population vector model." This model is based on the idea that neuronal coding can be analyzed in terms of a series of vectors, each based on specific movement parameters. In the present case, each vector gives the mean contribution of a population of muscle spindle afferents within one directionally tuned muscle. A given population vector points in the "preferred sensory direction" of the muscle to which it corresponds, and its length is the mean frequency of all the afferents within that muscle. Our working hypothesis was that the sum of these weighted vectors points in the same direction as the ongoing movement. The results show that each muscle spindle afferent, and likewise each muscle, has a specific preferred sensory direction, as well as a preferred sensory sector within which it is capable of sending sensory information to the central nervous system. Interestingly, the results also demonstrate that the preferred directions are the same as the directions of vibration-induced illusions. In addition, the results show that the neuronal population vector model describes the multipopulation proprioceptive coding of spatially oriented 2D limb movements, even at the peripheral sensory level, based on the sum vectors calculated from all the muscles involved in the movement. In an accompanying paper, the coding of more complex 2D movements such as those involved in drawing rectilinear and curvilinear geometrical shapes was investigated.

Proprioceptive population coding of two-dimensional limb movements in humans: II. Muscle-spindle feedback during "drawing-like" movements.

It was proposed to study the proprioceptive sensory coding of movement trajectories during the performance of two-dimensional "drawing-like" movements imposed on the tip of the foot. For this purpose, the activity of the muscle-spindle afferents from the Extensor digitorum longus, Tibialis anterior, Extensor hallucis longus, and Peroneus lateralis muscles was recorded from the lateral peroneal nerve using the microneurographic technique. The drawing movements, describing geometrical shapes such as squares, triangles, ellipses, and circles, were imposed at a constant velocity in both the clockwise and counterclockwise directions. A total number of 44 muscle-spindle afferents were tested, 36 of which were identified as primary and eight as secondary afferents. Whatever the shape of the imposed foot movement, the primary endings from one muscle never discharged throughout the whole trajectory (on average, they discharged for only 49.2% of the length of the trajectory), whereas all the secondary endings discharged for most part of the drawing trajectories (average: 84.8%). The relationship between afferent discharge rate and direction could be described with a cosine-shaped tuning function. The peak of this function corresponded to the preferred sensory direction of the receptor-bearing muscles. The whole path of a given geometrical drawing movement was found to be coded in turn by each of the primary afferents originating from each of the muscles successively stretched. The contribution of each population of muscle afferents from each ankle muscle was represented by a "population vector", whose orientation was the preferred direction of the muscle under consideration and whose length was the mean instantaneous frequency of the afferent population. The "sum vector" corresponding to the sum of all these weighted "population vectors" was found to point in the instantaneous direction of the drawing trajectory, i.e., the tangent to the trajectory. These findings suggest that trajectory information is already encoded at the peripheral level on the basis of the integrated inputs provided by sets of receptors belonging to all the muscles acting on a given joint.

Bradykinin and muscle stretch alter contralateral cat neck muscle spindle output.

The objective of this study was to investigate the activity of primary and secondary muscle spindle afferents (MSAs) in neck muscles, when the contralateral splenius or trapezius muscles were (1) exposed to i.m. injection of bradykinin (BK) (6-86 microg/ml), (2) stretched, (3) stretched during exposure to BK or (4) stretched after exposure to BK. It was found that injection of BK, muscle stretch and the combination of the two stimuli significantly excited primary and secondary MSAs via reflex effects onto static fusimotor neurones. BK, alone, and in combination with muscle stretch, induced more frequent, potent and long lasting effects as compared to muscle stretch. The effects of muscle stretch was significantly increased after exposure of BK, indicating that stretch-sensitive nerve-endings within neck muscles are sensitised by BK. The results are discussed in relation to the disturbances in motor co-ordination and proprioception found in patients suffering from chronic muscle pain in the cervical region.

The information transmitted by ensembles of primary spindle afferents is diminished when ketamine is used as a pre-anaesthetic.

The effect of pre-anaesthetic ketamine on ensemble coding of different stimuli consisting of muscle stretches of various amplitudes was studied for ensembles of simultaneously recorded primary muscle spindle afferents (MSAs). The experiments were conducted on 8 alpha-chloralose anaesthetised cats. Three of the cats received a pre-anaesthetic dose of ketamine (25 mg/kg) injected subcutaneously (ketamine group), while the remaining five animals did not (non-ketamine group). Data for ensemble coding were collected both before and after cutting the ventral root. A method based on principal component analysis and algorithms was used to quantify stimulus discrimination and an ANOVA tested differences between groups as well as differences due to ventral root cutting. When the fusimotor supply was intact, a general trend of an increase in the ability to discriminate stimuli with increasing ensemble size was observed for both groups, however, this trend was significantly greater for the non-ketamine group as compared to the ketamine group. When the ventral root was cut, the discrimination pattern for the non-ketamine group decreased significantly (as compared to before ventral root cutting), however, no change occurred for the ketamine group. Consequently, no difference in discrimination pattern was detected between groups after ventral root cutting. The reduction in information transmitted by ensembles of primary MSAs when ketamine is used as a pre-anaesthetic may suggest that ketamine elicits an adverse affect on the fusimotor system.

Alterations in information transmission in ensembles of primary muscle spindle afferents after muscle fatigue in heteronymous muscle.

This study showed that fatigue of the ipsilateral medial gastrocnemius muscle caused a clear-cut reduction in the ability of ensembles of primary muscle spindle afferents from the lateral gastrocnemius muscle to discriminate between muscle stretches of varying amplitude. The results were probably caused by reflex-mediated effects from chemosensitive group III and IV afferents onto the gamma-motoneurons projecting to lateral gastrocnemius muscle spindles. The experiments were conducted on seven cats anaesthetized with alpha-chloralose and a total of 41 primary muscle spindle afferents from the lateral gastrocnemius were registered. Afferents were simultaneously recorded in ensembles of three to 10 afferents. A method based on principal component analysis and algorithms for quantification of stimulus discrimination in ensembles of muscle afferents was used prior to, immediately following and five or more minutes after muscle fatigue had been induced to the ipsilateral medial gastrocnemius muscle. It is well established that the primary muscle spindle afferents play an important role in proprioception and kinaesthesia. Therefore the decrease in the accuracy of the information transmitted by ensembles of primary muscle spindle afferents caused by fatigue in an ipsilateral muscle implies concomitant effects on proprioception and kinaesthesia.

Ensemble coding of muscle stretches in afferent populations containing different types of muscle afferents.

Ensemble coding of simple mechanical stimuli (small sinusoidal stretches) was studied in simultaneously recorded mixed ensembles of primary- and secondary muscle spindle afferents (MSAs), and Golgi tendon organ (GTO) afferents recorded from L7-S1 dorsal root filaments. The experiments were made on 48 recorded afferents (29 primary MSAs, 6 secondary MSAs and 13 GTO afferents) in chloralose anaesthetised cats. For the analyses, we used a combination of principal component analysis and algorithms for quantification of stimulus discrimination. Mixed ensembles of primary- and secondary MSAs, and GTO afferents, discriminated significantly better between different muscle stretches than ensembles of only one or two types of these afferents. All kinds of ensembles showed a successive increase in discriminative ability with increased ensemble size and this ability seemed to level at larger populations. However, the increase in discriminative ability was significantly greater for the mixed ensembles. It is hypothesised that the main reason for the greater discriminative ability achieved by mixed ensembles, might be that the variation in response profiles (sensitivity tuning) among the individual afferents of the mixed ensemble will be larger than that for ensembles of only one type of afferent. Finally, the results in the present study give experimental support to some of the teleological arguments in favour of the ensemble coding theory.

A neural network approach to real-time spike discrimination during simultaneous recording from several multi-unit nerve filaments.

A multi-channel, real-time, unsupervised spike discriminator was developed in order to reconstruct single spike trains from several simultaneously recorded multi-unit nerve filaments. The program uses a Self Organising Map (SOM) algorithm for the classification of the spikes. In contrast to previous similar techniques, the described method is made for use on a PC, and the method may thus be implemented at relatively low cost. In order to test the accuracy of the program, a robustness test was performed, where noise with different RMS levels was superimposed on the spikes. Furthermore, the maximal classification rate was determined. The program is easy to use, since the only manual inputs needed are the voltage threshold for spike detection, and the number of units present in each recorded nerve filament.

A method for analysis of encoding of stimulus separation in ensembles of afferents.

This paper describes a new method for the analysis of ensemble coding in populations of receptor afferents. The method is based on principal component analysis (PCA) combined with algorithms for calculation of stimulus separation (discrimination). It allows a quantification of the ability of ensembles of afferents to discriminate between stimuli of different intensity, and it can also be used to calculate to which extent each afferent in an ensemble contributes to the discrimination. Since the emphasis is put on discrimination of stimuli rather than on similarities between the representations of the stimulus in the ensemble response and the stimulus itself, it involves no implicit assumption about the unknown decoding mechanisms in the CNS. Examples of results obtained with analysis of and comparison of simultaneously recorded and pooled sequentially recorded populations of primary muscle spindle afferents are given. These results support some of the general arguments in favour of the ensemble coding theory, and indicate that the results of studies made on populations of simultaneously recorded receptor afferents may differ considerably from results obtained in studies on pooled sequentially recorded afferents. In contrast to when ensemble responses are based on parallel recordings, population responses constructed from sequential recordings will be distorted as a result of unavoidable temporal variations in physiological conditions.

The role of the gamma-system for improving information transmission in populations of Ia afferents.

Ensemble coding of simple mechanical stimuli (small sinusoidal stretches) was studied in populations of simultaneously recorded primary muscle spindle afferents (MSAs). The experiments were made on 39 primary MSAs in chloralose anaesthetised cats. For the analyses we used a combination of principal component analysis and algorithms for quantification of stimulus discrimination. Ensembles of primary MSAs discriminated better between different muscle stretches than individuals, and showed a successive increase in discriminative ability with increasing ensemble size. The ensemble effect disappeared after cutting the ventral roots, indicating an important role for the fusimotor system in ensemble coding. Simultaneously recorded ensembles of MSAs showed significantly better discriminative ability than pooled sequentially recorded MSAs. This difference was abolished by the cutting of the ventral roots. It is hypothesised that, since the muscle spindles are connected to each other via secondary MSAs (projecting to gamma-motoneurones to other spindles), the fusimotor-muscle spindle system may constitute a neural network, in which each 'neuron' (i.e., each individual muscle spindle) is influenced by the activity in the whole network. In populations of pooled sequentially recorded MSAs, the connections would not exist. Thus, the population would lose its neural network quality, and the encoding ability of the population would decrease.

Influences on the gamma-muscle spindle system from muscle afferents stimulated by increased intramuscular concentrations of bradykinin and 5-HT.

There is evidence that static muscular contractions induce a release of bradykinin (BK) in the working muscle, and that increased concentration of BK and 5-HT in a muscle increases the discharge rate of a subpopulation of group III and group IV muscular afferents. It is also known that activity in group III and IV muscle afferents may activate gamma-motoneurones to both homonymous and heteronymous muscles. The aim of the present study was to investigate whether increased concentration of BK and 5-HT in one muscle may influence the activity in primary and secondary muscle spindle afferents (MSAs) from the chemically affected muscle and from surrounding muscles, via fusimotor reflexes. The experiments were made on six cats anaesthetised with alpha-chloralose. The triceps surae (GS) and the posterior biceps and semitendinosus (PBSt) muscles were subjected to sinusoidal stretches. Simultaneous recordings of 2-11 MSAs from these muscles were made and the mean rate of firing and the modulation for each MSA were determined. Responses of 47 MSAs (26 PBSt and 21 GS) were recorded. The responsiveness of the MSAs to injections of BK (9-100 mg/ml, 0.5-1.0 ml) and 5-HT (25-150 mg/ml, 0.5-1.0 ml) was 89% and 83%, respectively, for injections into the arterial supply of the ipsilateral GS muscle, and 84% and 40% respectively for injections to the contralateral GS muscle. Of 10 secondarLy MSAs, only one was unresponsive to BK injections, while several MSAs responded to both ipsilateral and contralateral BK injections.(ABSTRACT TRUNCATED AT 250 WORDS)

Influences on the gamma-muscle-spindle system from contralateral muscle afferents stimulated by KCl and lactic acid.

The aim of the present study was to investigate whether increased concentrations of lactic acid and potassium chloride (KCl) in contralateral muscles can influence the sensitivity of primary and secondary muscle spindle afferents (MSAs) from ipsilateral extensor and flexor muscles. The experiments were performed on 7 cats anaesthetised with alpha-chloralose. Recordings were made simultaneously from 2-12 single MSAs from the triceps surae (GS) and/or the posterior biceps and semitendinosus muscles (PBSt). The mean rate of firing and the amplitude of a fitted sine curve of MSA responses to sinusoidal stretching of the receptor bearing muscles were determined. Responses of 42 primary MSAs (17 from PBSt and 25 from GS) were recorded. On 33 of these, clear-cut alterations in sinusoidal response were evoked by injection of 1 ml KCl (200-400 mM) or 1 ml lactic acid (20-50 mM) into the arterial supply of the contralateral GS or PBSt muscles. Six out of 8 secondary MSAs showed sizeable effects to increased intramuscular concentrations of KCl and/or lactic acid (3 from PBSt and 3 from GS). On both primary and secondary MSAs, from GS as well as from PBSt muscles, the large majority of effects were excitatory. All effects on secondary MSAs were compatible with reflex actions on static fusimotor neurones, whereas on primary MSAs different types of reflex responses were observed (i.e. pure static, pure dynamic and mixtures of static and dynamic fusimotor actions). Stimuli related alterations in MSA responses were completely abolished when the contralateral GS or PBSt nerves were anaesthetised.(ABSTRACT TRUNCATED AT 250 WORDS)

Influences on the gamma-muscle-spindle system from muscle afferents stimulated by increased intramuscular concentrations of arachidonic acid.

There is evidence that static muscular contractions induce a release of arachidonic acid (AA) in the working muscle and that increased concentration of AA in a muscle increases the discharge rate of a subpopulation of groups III and IV muscular afferents. It is also known that activity in groups III and IV muscle afferents may activate gamma-motoneurones to both homo- and heteronymous muscles. The aim of the present study was to investigate if increased concentration of AA in one muscle may influence the activity in primary and secondary muscle spindle afferents (MSAs) from the chemically affected muscle and from surrounding muscles, via fusimotor reflexes. The experiments were made on five cats anaesthetized with alpha-chloralose. The triceps surae (GS) and the posterior biceps and semitendinosus (PBSt) muscles were subjected to sinusoidal stretches. Simultaneous recordings of 2-12 MSAs from these muscles were made and the mean rate of firing and the modulation for each MSA were determined. Responses of 36 MSAs (17 PBSt and 19 GS) were recorded. The responsiveness of the MSAs to injections of AA (0.3-1.0 mg; 0.3-1 ml) was 86% (n = 36) for injections into the arterial supply of the ipsilateral GS muscle and 45% (n = 20) for injections to the contralateral GS muscle. Out of 14 secondary MSAs, only one was unresponsive to ipsilateral AA injections while two of eight were unresponsive to contralateral AA injection. The majority of responses were compatible with predominantly static or mixed dynamic and static fusimotor activation. None of the effects were compatible with inhibition of fusimotor activity. The duration of the effects were usually 2-4 min. However, on some occasions the elevations in MSA activity persisted for up to 1 h. Local anaesthesia of the nerve to the injected muscle always abolished the effects of the injections and control injections of the solution in which the AA was dissolved were ineffective in changing the MSA responses. I.v. injections occasionally induced effects on the MSAs, but such effects were significantly different from those caused by close arterial muscle injections. Thus, increased concentration of AA may excite primary and secondary MSAs from homo- as well as heteronymous muscles, including contralateral muscles, most probably via fusimotor reflexes evoked by activity in chemosensitive muscle afferents.

A multichannel hook electrode for simultaneous recording of up to 12 nerve filaments.

This paper describes a simple multichannel hook electrode specially adapted for simultaneous recording from several dorsal root filaments in the cat. The electrode consists of a 3-mm-thick PVC plate and accommodates 12 silver wire hooks placed in notches in the plate. It has a semicircular shape, and the notches are milled radially in the plate. With this electrode we have made successful recordings of single-unit activity from 12 muscle afferents for periods of up to 8 h. In contrast to more rigid and complicated devices, e.g., developed to assess activity of populations of individual neurones from single-channel recordings of multi-unit activity, it also permits selection of the types of afferents to be recorded. The use of this simple device and an appropriate number of amplifiers and window discriminators can greatly enhance the quality of studies on populations of afferents in animals and may also decrease the number of laboratory animals needed.