Comparison of the effects of two different handling methods on the subsequent behaviour of Anglo-Arabian foals toward humans and handling

Tactile contact is often used to improve the human–foal relationship, in particular during sensitive periods such as weaning. However, the method used to provide the contact (forced or unforced) may affect subsequent reactions to humans. The aim of our study was to compare the effect of forced and unforced handling at weaning on horses’ behaviour towards humans and handling.A total of 23 Anglo-Arabian foals received individual human contact at weaning. Constrained-handled foals (forced human contact; FC, n = 8) were stroked while being restrained (i.e., held by the halter without being led) so that human contact was forced. Unconstrained-handled foals (unforced human contact; UC, n = 7) were stroked without any restraint and could thus avoid contact if they so wanted. Each individual was handled 5 min twice a day for 14 consecutive days. Control foals were not handled (no contact; NC, n = 8). Foals’ reactions towards a human (e.g. proximity and contact seeking), and their manageability (fitting of a halter) were then assessed. The animals from the different treatments were then mixed and tested again 4 months later. Non-parametric Mann–Whitney U-tests were used to compare experimental groups.Only forced human contact reduced fear reactions toward humans (e.g. shorter latency to approach a passive human, P < 0.01; shorter latency to touch the foal, P < 0.05) in the short-term (immediately after the handling sessions) and increased foals’ manageability in a familiar environment (shorter latency to be placed a halter on and less defences displayed, P < 0.05). It had no effect in an unfamiliar environment. These effects did not last longer than 4 months.Forced human contact thus appears to be more efficient in improving human–horse relationships and subsequent handling than unforced human contact. However, the lack of effect on reactivity towards humans in an unfamiliar environment, and the lack of long-term effects suggest that this method needs to be improved to be more effective. Some of our results suggest that horses could perceive human contact (i.e., stroking) as positive, however further studies are needed to confirm this hypothesis.     

Tactile contact is required for early estrogen treatment to alter the sexual partner preference of female zebra finches

Sex steroid actions during early development appear to play a role in the development of sex differences in sexual partner preference (SPP, preference for males vs. females) in several species of mammals and in the socially monogamous pair bonding zebra finch (Taeniopygia guttata). Female finches treated with estrogen as nestlings exhibit varying degrees of masculinized SPP as adults, but only if they have been housed in all-female groups during the juvenile and young adult period, suggesting that the estrogen effect may involve social experience and possibly sexual imprinting. Because tactile contact is important for consolidation of imprinted preferences in this species, it was predicted that early estrogen treatment would alter preferences of females only if they were allowed to have tactile contact with other females. Subjects were injected with estradiol benzoate or with oil (normal controls) daily for the first 2 weeks post-hatching. At age 45 days, they were housed in a mixed sex aviary (normal controls), in an all-female aviary allowing tactile contact (group EB-TC), or in an all-female aviary with no tactile contact (group EB-NTC). At 100+ days, birds were given two-choice SPP tests followed by aviary tests of SPP. EB-TC females did not show the sex-typical preference for male stimuli, and differed significantly from the controls on several measures. EB-NTC females preferred males and never differed significantly from controls. These results show that tactile contact after age 45 days is essential for an EB effect on SPP, supporting the hypothesis that hormones and sexual imprinting together contribute to SPP.     

Elicited activity of the sensorimotor cortex after chronic unilateral exclusion of the spinocervical tract

Elicited potentials of the sensorimotor cortex were investigated in chronic experiments before and after unilateral injury to the spinocervical tract at the C₃ level. Such injury led to a considerable disturbance of tactile and proprioceptive reactions but did not lower the amplitude of potentials elicited in the cerebral cortex by irritation of a limb on the injured side. On the intact side, there was an increase in the amplitude of the early response components to irritation of the extremity on this side. In 1–2 months after the operation proprioceptive sensitivity and motor activity had returned to normal, but the reactions to tactile irritation failed to reappear. Apparently the presence of a high level of afferent input into the cerebral cortex is insufficient for retention of somatic sensitivity. It is suggested that an inflow over several channels plays a role in providing the spatial-temporal sequence required for activating cortical neurons.Institute of Higher Nervous Activity and Neurophysiology, Academy of Sciences, USSR, Moscow. Translated from Neirofiziologiya, Vol. 2, No. 5, pp. 469–474, September–October, 1970.     

The Way Humans Behave Modulates the Emotional State of Piglets

The emotional state can influence decision-making under ambiguity. Cognitive bias tests (CBT) proved to be a promising indicator of the affective valence of animals in a context of farm animal welfare. Although it is well-known that humans can influence the intensity of fear and reactions of animals, research on cognitive bias often focusses on housing and management conditions and neglects the role of humans on emotional states of animals. The present study aimed at investigating whether humans can modulate the emotional state of weaned piglets. Fifty-four piglets received a chronic experience with humans: gentle (GEN), rough (ROU) or minimal contact (MIN). Simultaneously, they were individually trained on a go/no-go task to discriminate a positive auditory cue, associated with food reward in a trough, from a negative one, associated with punishments (e.g. water spray). Independently of the treatment (P = 0.82), 59% of piglets completed the training. Successfully trained piglets were then subjected to CBT, including ambiguous cues in presence or absence of a human observer. As hypothesized, GEN piglets showed a positive judgement bias, as shown by their higher percentage of go responses following an ambiguous cue compared to ROU (P = 0.03) and MIN (P = 0.02) piglets, whereas ROU and MIN piglets did not differ (P > 0.10). The presence of an observer during CBT did not modulate the percentage of go responses following an ambiguous cue (P > 0.10). However, regardless of the treatment, piglets spent less time in contact with the trough following positive cues during CBT in which the observer was present than absent (P < 0.0001). This study originally demonstrates that the nature of a chronic experience with humans can induce a judgement bias indicating that the emotional state of farm animals such as piglets can be affected by the way humans interact with them.     

The Way to a Man's Heart Is through His Stomach: What about Horses?

Background

How do we bond to one another? While in some species, like humans, physical contact plays a role in the process of attachment, it has been suggested that tactile contact''s value may greatly differ according to the species considered. Nevertheless, grooming is often considered as a pleasurable experience for domestic animals, even though scientific data is lacking. On another hand, food seems to be involved in the creation of most relationships in a variety of species.

Methodology/Principal Findings

In this study, we used the horse training context to test the effects of food versus grooming during repeated human-horse interactions. The results reveal that food certainly holds a key role in the attachment process, while tactile contact was here clearly insufficient for bonding to occur.

Conclusion/Significance

This study raises important questions on the way tactile contact is perceived, and shows that large inter-species differences are to be expected.     

Tactile sensitivity of normal and autistic children

Many children with autistic spectrum disorders have unusual reactions to certain sensory stimuli. These reactions vary along a hyper- to hypo-responsivity continuum. For example, some children overreact to weak sensory input, but others do not respond negatively to even strong stimuli. It is typically assumed that this deviant responsivity is linked to sensitivity, although the particular stage of sensory processing affected is not known. Psychophysical vibrotactile thresholds of six male children (age: 8–12) who were diagnosed to have autistic spectrum disorders and six normal male children (age: 7–11) were measured by using a two-alternative forced-choice task. The tactile stimuli were sinusoidal displacements and they were applied on the terminal phalanx of the left middle finger of each subject. By using a forward-masking paradigm, 40- and 250-Hz thresholds of the Pacinian tactile channel and 40-Hz threshold of the Non-Pacinian I tactile channel were determined. There was no significant difference between the thresholds of autistic and normal children, and the autistic children had the same detection and masking mechanisms as the normal children. The sensory responsivity of each subject was tested by clinical questionnaires, which showed again no difference between the two subject groups. Furthermore, no significant correlations could be found between the questionnaire data and the psychophysical thresholds. However, there was a high correlation between the data from the tactile and emotional subsets of the questionnaires. These results support the hypothesis that the hyper- and hypo-responsivity to touch, which is sometimes observed in autistic spectrum disorders, is not a perceptual sensory problem, but may probably be emotional in origin.     

触觉剥夺后豚鼠桶状皮质DCX 阳性细胞的实验研究*

目的:观察豚鼠单侧触觉剥夺后豚鼠两侧桶状皮质的DCX阳性细胞数量的差别,探讨触觉剥夺对豚鼠桶状皮质神经发生的影响。方法:12只健康豚鼠随机分为2组,每组6只,制作豚鼠单侧(右侧)触觉(胡须)剥夺模型,之后常规饲养1月和2月灌注取材,用免疫组化和免疫荧光双标法观察同一只豚鼠两侧桶状皮质的DCX阳性细胞情况并比较其数量差异。结果:DCX免疫组织化学染色显示两实验组大脑皮层barrel区DCX阳性细胞数实验侧均明显多于对照侧;NeuN免疫组织化学染色显示两实验组动物大脑皮层barrel区两侧NeuN阳性细胞数差别无统计学意义;DCX和NeuN免疫荧光双标染色显示两实验组大脑皮层barrel区均可见双标细胞存在。结论:触觉剥夺后豚鼠两侧桶状皮质的DCX阳性细胞数具有明显差异性,可能是神经再生的表现。     

Tactile sensitivity of normal and autistic children

Many children with autistic spectrum disorders have unusual reactions to certain sensory stimuli. These reactions vary along a hyper- to hypo-responsivity continuum. For example, some children overreact to weak sensory input, but others do not respond negatively to even strong stimuli. It is typically assumed that this deviant responsivity is linked to sensitivity, although the particular stage of sensory processing affected is not known. Psychophysical vibrotactile thresholds of six male children (age: 8-12) who were diagnosed to have autistic spectrum disorders and six normal male children (age: 7-11) were measured by using a two-alternative forced-choice task. The tactile stimuli were sinusoidal displacements and they were applied on the terminal phalanx of the left middle finger of each subject. By using a forward-masking paradigm, 40- and 250-Hz thresholds of the Pacinian tactile channel and 40-Hz threshold of the Non-Pacinian I tactile channel were determined. There was no significant difference between the thresholds of autistic and normal children, and the autistic children had the same detection and masking mechanisms as the normal children. The sensory responsivity of each subject was tested by clinical questionnaires, which showed again no difference between the two subject groups. Furthermore, no significant correlations could be found between the questionnaire data and the psychophysical thresholds. However, there was a high correlation between the data from the tactile and emotional subsets of the questionnaires. These results support the hypothesis that the hyper- and hypo-responsivity to touch, which is sometimes observed in autistic spectrum disorders, is not a perceptual sensory problem, but may probably be emotional in origin.     

Extracellular matrix changes in early osteochondrotic defects in foals: a key role for collagen?

Osteochondrosis (OC) is the most important developmental orthopaedic disease in the horse. Despite some decades of research, much of the pathogenesis of the disorder remains obscure. Increasing knowledge of articular cartilage development in juvenile animals led to the presumption that the role of collagen in OC might be more important than previously thought. To study collagen characteristics of both cartilage and subchondral bone in young (5 and 11 months of age) horses, samples were taken of subchondral bone and articular cartilage from a group of 43 Dutch Warmblood foals and yearlings that suffered from varying degrees of OC. Based on a histological classification, lesions were graded as early, middle and end stage. Collagen content and some posttranslational modifications (lysyl hydroxylation, hydroxylysylpyridinoline (HP) and lysylpyridinoline (LP) cross-links) were determined, as was proteoglycan content. Data were compensated for site effects and analysed for differences due to the stage of the lesion. In early lesions total collagen was significantly decreased in both cartilage and subchondral bone of 5- and 11-month-old foals. Also in cartilage, HP cross-linking was reduced in the early lesions of 5- and 11-month-old foals, while LP cross-linking was decreased in subchondral bone of the end-stage lesions of both 5- and 11-month-old foals. Hydroxylysine content was unaffected. Collagen content remained reduced in cartilage from middle- and end-stage lesions, but returned to normal in subchondral bone. In cartilage there was a decrease in proteoglycan content in the end-stage lesions of both age groups. Thus, alterations of the collagen component, but not of the proteoglycan component, of the extracellular matrix might play a role in early OC. More severe lesions show a more general picture of an unspecific repair reaction. Biomarkers of collagen metabolism can be expected to be good candidates for early detection of OC.     

Implicit Attitudes towards People with Intellectual Disabilities: Their Relationship with Explicit Attitudes,Social Distance,Emotions and Contact

Implicit attitude research has expanded rapidly over the last decade and is seen as very promising as it counters biases present in much attitude research such as social desirability. However, most research in the area of intellectual disabilities has focused on explicit attitudes alone. This study examined implicit attitudes to this population and also examined their association with emotional reactions and contact, which have previously been found to have a significant influence on attitudes and stigma. A web based survey consisting of a single target Implicit Association Test, measures of explicit attitudes, social distance, and emotional reactions towards and contact with individuals with intellectual disabilities was completed by 326 adult UK residents. Implicit attitudes were not significantly associated with explicit attitudes, social distance or emotional reactions. Instead there were small to moderate associations between emotional reactions and explicit attitudes and social distance. Implicit attitudes did not vary according to participants’ level of contact with individuals with intellectual disabilities, type of the contact relationship (voluntary versus involuntary), gender or educational attainment. In contrast, these participant characteristics did affect explicit attitudes and social distance. Implicit attitudes towards individuals with intellectual disabilities were somewhat negative and, unlike explicit attitudes and stigma, did not vary according to participant demographics or contact. As they may have a negative impact on the lives of people with intellectual disabilities, implicit attitudes merit increased attention in research and interventions in the intellectual disabilities field.     

Emotion regulation and touch in infants: the role of cholecystokinin and opioids

Behavioral-pharmacological research in infant rats supports the role of cholecystokinin (CCK) and opioid peptides in mediating early learning of new associations with aspects of the nest and dam, such as maternal odor, milk, and contact. The current paper reviews research that examines the hypothesis that these neuropeptide systems are further involved in mediating emotion regulation in infants, thus playing a role in the emergence of stress-reactivity and other motivational systems. The beneficial effects of maternal proximity, handling, and touch on the development of emotion regulation have been demonstrated in both human and animal models. Interventions that promote tactile stimulation of the infant ("touch therapy") and infant-mother contact ("skin-to-skin contact" or "kangaroo care") have been shown to improve the infant's ability to self-regulate, and to moderate the effects of some risk factors. Theoretical perspectives and empirical findings regarding emotion regulation in infants are first discussed. This is followed by a review of work providing evidence in animal models (and suggestive evidence in humans) for the importance of CCK and opioid neuropeptides in affecting infant emotion regulation and the impact of touch-based interventions, in particular in the context of infant-mother attraction, contact, separation, and attachment.     

Characteristics of cryptic/ectopic and contralateral scrotal testes in dogs between 1 and 2 years of age

Testicular malposition represents a common developmental genital defect in dogs and can affect one or both testes. In both humans and dogs, unilateral cryptorchism is more frequently detected and thought to be the expression of a genetic abnormality affecting both the undescended and scrotal testis. In the dog, there is evidence of degenerative processes affecting the maldescended testis. However, the histologic and functional changes that occur in the scrotal testis of unilateral cryptorchid or ectopic individuals remain a source of debate. Because the bilateral surgical removal of the testes leads to some undesirable side effects, the aim of this study was to evaluate the necessity for performing bilateral orchiectomy in young unilateral cryptorchid dogs. A morphologic study of both cryptic/ectopic and scrotal testes in young dogs affected by unilateral testicular maldescent was therefore conducted. The study was conducted on 10 dogs aged 1 to 2 yr and affected by unilateral testicular maldescent. We found that, in young dogs, even if no neoplastic lesions were observed, morphologic abnormalities are detectable between 1 and 2 yr of age in the maldescended testes with severity dependent on testicular position. In contrast, in the scrotal testes, the histologic and immunohistochemical exam failed to find signs of incorrect development or morphologic abnormalities. The results seem to suggest that, though the early removal of the undescended testis is recommended, continuous monitoring of the scrotal testis for the life of the dog is preferable to removing it considering the undesirable side effects related to castration.     

Facial disfigurement is treated like an infectious disease

The behavioral avoidance of people with facial disfigurement is well documented, but its psychological basis is poorly understood. Based upon a disease avoidance account of stigmatization, we conducted the first empirical test of whether facial disfigurement—naevus flammeus (a port wine stain) here—can trigger the same set of emotional and behavioral responses as a contagious disease (influenza). Ninety-eight participants contacted props, which they had seen used either by a healthy confederate or by a confederate simulating medical conditions affecting the face—birthmark and influenza. Behavioral avoidance (e.g., willingness to handle the prop) and facial display of disgust were recorded across five levels of prop contact varying from no contact to contact with the mouth. Behavioral avoidance and disgust displays, especially with oral contact, were equivalent in the birthmark and influenza conditions, with both significantly exceeding reactions to the healthy confederate. These results support the theory that humans have an evolved predisposition to avoid individuals with disease signs, which is mediated by the emotion of disgust. This implicit avoidance occurs even when they know explicitly that such signs—the birthmark here—result from a noncontagious condition.     

新生期触觉刺激和母婴分离对雌性大鼠成年后的焦虑和情绪记忆的影响

采用split-litter法对仔鼠进行分组和处理,共5组NTS组(未经实验人员抓握和标记),PND2—9TS组和PND10—17TS组(分别在仔鼠出生后的2—9天、10—17天,每天短暂抓握和标记仔鼠),PND2—9MS组和PND10—17MS组(分别在仔鼠出生后的2—9天、10—17天,除了按TS组相同方式抓握并在不同部位标记外,每天把仔鼠与母鼠分离1h)。待雌鼠成年后,进行明/暗箱测试和一次性被动回避反应测试。结果发现与NTS组相比,PND2—9TS组和PND10—17TS组的雌鼠在明/暗箱测试中停留于明室的累计时间明显较长,在被动回避作业中的重测试潜伏期也明显较长,表明新生期的触觉刺激经历减少雌性大鼠成年后在新异环境中的焦虑,并改善情绪记忆。与相应TS组相比,MS处理组的所有行为指标都无显著性差异,说明短时间母婴分离对雌鼠成年后的焦虑和情绪记忆无明显影响。结果提示,新生期的触觉刺激和母婴分离经历对仔鼠神经系统的发育产生不同的长期效应。     

Recruitment of occipital cortex during sensory substitution training linked to subjective experience of seeing in people with blindness

Over three months of intensive training with a tactile stimulation device, 18 blind and 10 blindfolded seeing subjects improved in their ability to identify geometric figures by touch. Seven blind subjects spontaneously reported 'visual qualia', the subjective sensation of seeing flashes of light congruent with tactile stimuli. In the latter subjects tactile stimulation evoked activation of occipital cortex on electroencephalography (EEG). None of the blind subjects who failed to experience visual qualia, despite identical tactile stimulation training, showed EEG recruitment of occipital cortex. None of the blindfolded seeing humans reported visual-like sensations during tactile stimulation. These findings support the notion that the conscious experience of seeing is linked to the activation of occipital brain regions in people with blindness. Moreover, the findings indicate that provision of visual information can be achieved through non-visual sensory modalities which may help to minimize the disability of blind individuals, affording them some degree of object recognition and navigation aid.     

Enhanced tactile performance at the destination of an upcoming saccade

Previous work has demonstrated that upcoming saccades influence visual and auditory performance even for stimuli presented before the saccade is executed. These studies suggest a close relationship between saccade generation and visual/auditory attention. Furthermore, they provide support for Rizzolatti et al.'s premotor model of attention, which suggests that the same circuits involved in motor programming are also responsible for shifts in covert orienting (shifting attention without moving the eyes or changing posture). In a series of experiments, we demonstrate that saccade programming also affects tactile perception. Participants made speeded saccades to the left and right side as well as tactile discriminations of up versus down. The first experiment demonstrates that participants were reliably faster at responding to tactile stimuli near the location of upcoming saccades. In our second experiment, we had the subjects cross their hands and demonstrated that the effect occurs in visual space (rather than the early representations of touch). In our third experiment, the tactile events usually occurred on the opposite side of upcoming eye movement. We found that the benefit at the saccade target location vanished, suggesting that this shift is not obligatory but that it may be vetoed on the basis of expectation.     

Preserved functional specialization for spatial processing in the middle occipital gyrus of the early blind

The occipital cortex (OC) of early-blind humans is activated during various nonvisual perceptual and cognitive tasks, but little is known about its modular organization. Using functional MRI we tested whether processing of auditory versus tactile and spatial versus nonspatial information was dissociated in the OC of the early blind. No modality-specific OC activation was observed. However, the right middle occipital gyrus (MOG) showed a preference for spatial over nonspatial processing of both auditory and tactile stimuli. Furthermore, MOG activity was correlated with accuracy of individual sound localization performance. In sighted controls, most of extrastriate OC, including the MOG, was deactivated during auditory and tactile conditions, but the right MOG was more activated during spatial than nonspatial visual tasks. Thus, although the sensory modalities driving the neurons in the reorganized OC of blind individuals are altered, the functional specialization of extrastriate cortex is retained regardless of visual experience.     

Changing reference frames during the encoding of tactile events

The mindless act of swatting a mosquito on the hand poses a remarkable challenge for the brain. Given that the primary somatosensory cortex maps skin location independently of arm posture [1, 2], the brain must realign tactile coordinates in order to locate the origin of the stimuli in extrapersonal space. Previous studies have highlighted the behavioral relevance of such an external mapping of touch, which results from combining somatosensory input with proprioceptive and visual cues about body posture [3-7]. However, despite the widely held assumption about the existence of this remapping process from somatotopic to external space and various findings indirectly suggesting its consequences [8-11], a demonstration of its changing time course and nature was lacking. We examined the temporal course of this multisensory interaction and its implications for tactile awareness in humans using a crossmodal cueing paradigm [12, 13]. What we show is that before tactile events are referred to external locations [12-15], a fleeting, unconscious image of the tactile sensation abiding to a somatotopic frame of reference rules performance. We propose that this early somatotopic "glimpse" arises from the initial feed-forward sweep of neural activity to the primary somatosensory cortex, whereas the later externally-based, conscious experience reflects the activity of a somatosensory network involving recurrent connections from association areas.     

The "Moi-peau"

To construct a coherent identity, humans must distinguish what belongs to the external, perceived world from what belongs to their own inner world and the inner world of other individuals. Based on the theory developed by S. Freud and on work by ethologists, a number of psychoanalysts (J. Bowlby, R.A. Spitz, D.W. Winnicott, etc.) have underlined the importance of early tactile exchanges with the mother if a child is to become an autonomous individual who feels secure within what he or she perceives to be sound and reliable mental and physical boundaries. More recently, other psychoanalysts (E. Bick, W.R. Bion, etc.) have studied the fantasized mental structures that form the limits between an individual's inner mental space and the external world (including other individuals). As part of this theoretical psychoanalytical movement, Didier Anzieu, a French psychoanalyst, started to develop the concept of the "Moi-peau" in 1974. The "Moi-peau" designates a fantasized reality that a child uses during its early development to represent itself as "me", based on its experience of the body surface. The child, enveloped in its mother's care, fantasizes of a skin shared with its mother: on one side the mother (the outer layer of the "Moi-peau"), and on the other side the child (the inner layer of the "Moi-peau"). These two layers must separate gradually if the child is to acquire its own me-skin. D. Anzieu's work allowed dermatologists and other specialists, such as pediatricians, to focus on the quality of early tactile exchanges between a mother and her child, including the child with a chronic skin disorder. It also helped dermatologists to recognize patients with "borderline" states, which are particularly frequent in dermatology (ereutophobia, dysmorphophobia, tattooing, self-mutilation, artefacta dermatitis). These borderline patients are adults who, as a result of their mental conflicts, adopt defense mechanisms derived from both neurotic and psychotic functioning. Their complaints reflect difficulties with the solidity of their mental and physical limits: their real skin is metaphorically linked to the fantasized mental structure that delimits the individual mental space. Among other clinical characteristics, they have a "pathology of action" and frequently attack their own skin, paradoxically, in order to test the solidity and reliability of their own limits. Finally, D. Anzieu's work encouraged dermatologists to use psychotherapeutic approaches in parallel to classical dermatologic approaches, when necessary, and helped them better understand how psychoanalysts work with patients who have not yet acquired their own "Moi-peau". As a result, D. Anzieu was among the first to reconcile dermatologists and psychoanalysts.     

Application of chemical selective cleavage methods to analyze post-translational modification in proteins

Three chemical specific cleavage reactions, one for the carboxyl side of aspartyl peptide bonds, one for the carboxyl side of asparaginyl peptide bonds and another for the amino side of seryl/threonyl peptide bonds have been recently established. Additionally, these reactions simultaneously react on several post-translationally modified groups in peptides or proteins. The modified groups cover the external modifications N-formyl, N-acetyl, N-pyroglutamyi residues and C-terminal-alpha amide, as well as the internal modifications such as O-acetyl serine, phosphorylated serine/tyrosine, sulfonylated tyrosine, glycosylated serine/threonine and glycosylated asparagine. These three cleavage reactions relate to key amino acids for modifications, deamidation for asparagine, phosphorylation and acetylation for serine, and glycosylation for asparagine, serine and threonine. The chemical reactions on these modifications change the peptide mapping pattern, and information from these reactions may contribute characterization and location of post-translational modified groups in the protein.