Delayed onset muscle soreness (DOMS) is a subacute pain state arising 24–48 hours after a bout of unaccustomed eccentric muscle contractions. Functional magnetic resonance imaging (fMRI) was used to examine the patterns of cortical activation arising during DOMS-related pain in the quadriceps muscle of healthy volunteers evoked by either voluntary contraction or physical stimulation. The painful movement or physical stimulation of the DOMS-affected thigh disclosed widespread activation in the primary somatosensory and motor  cortices, stretching far beyond the corresponding areas somatotopically related to contraction or physical stimulation of the thigh; activation also included a large area within the cingulate cortex encompassing posteroanterior regions and the cingulate motor area. Pain-related activations were also found in premotor (M2) areas, bilateral in the insular cortex and the thalamic nuclei. In contrast, movement of a DOMS-affected limb led also to activation in the ipsilateral anterior cerebellum, while DOMS-related pain evoked by physical stimulation devoid of limb movement did not. Functional magnetic resonance imaging (fMRI) has revealed several aspects on how acute and chronic pain is processed in the human brain. Most pain imaging studies predominantly focus on the investigation of cerebral processing of cutaneous pain in spite of the fact that muscle and/or musculoskeletal pain entail more common clinical problems and are also widespread conditions amongst athletes To study the central processing of muscle pain by fMRI in a more physiological context we used eccentric muscle exercise and subsequent painful delayed onset muscle soreness (DOMS). In unaccustomed subjects eccentric exercise frequently results in the formation of DOMS, a form of subacute muscle pain, which appears 24 to 72 hours after exercise. Clinically, DOMS is a common but self-limiting condition that requires no treatment. In the present study, we induced DOMS locally confined to the right-sided quadriceps muscle group. Our study is the first to use DOMS to study its pain-related cerebral activation pattern in an fMRI environment. Within the fMRI environment, we evoked DOMS-related pain by voluntary painful contraction, but also by externally applied physical stimulation in the form of localized pressure which was moved over the sore partitions of the muscle by using a custom-designed heavy marble roll. The latter paradigm allowed separation of DOMS-related pain from limb movement-related brain activation. Our study is the first to highlight the central pathophysiology and activation pattern of this very common form of muscle pain.The fMRI study included eight right-handed healthy volunteers , who were either students or physicians and already experienced in participating in psychophysical studies. All participants participated voluntarily in the experiments and none of them were taking medication that might have interfered with the induction of subacute muscle soreness or with pain sensations . The study adhered to the claims of the Declaration of Helsinki and the protocol was approved by the local ethics committee .DOMS is a physiological pain state that occurs following a bout of eccentric muscle contractions and peaks in intensity 24-48 hours after exercise The intensity of the pain was quantified and monitored using the visual analogue scale (VAS) ranging from 0 to 100% as previously described . A red light indicated the stimulation period, during which the subjects were asked to repeatedly contract their right (sore) quadriceps muscle. Indicating control condition, the light switched to green, and the subjects to their healthy, left quadriceps group, maintaining phasic contractions over the period of the non-painful stimulus (control condition). In detail, for the contraction-related task, the subjects were advised to contract their quadriceps muscle either with maximum strength or until maximal tolerable pain was perceived and then to relax and contract the muscle again. One cycle of contraction and subsequent relaxation resulted in approximately 3 seconds and was repeated during the 24 seconds of stimulation until the light switched off to indicate baseline condition. The second paradigm consisted in physical stimulation of the sore areas of the thigh by externally applied localized pressure. It was used to separate brain activation in response to deep muscle pain from activation resulting from muscle contraction. Due to the slow adaptation properties of sensitized nociceptors (in deep tissues), pain sensation in response to tonic locally applied pressure dampens within seconds Shortly before subjects attended the fMRI scanner they were instructed to refrain from movement as much as possible. In the scanner the subjects' head was immobilized by fixing the head binaurally with a thrust device. A mirror was mounted above the eyes to allow the subject to see the light indicating the protocol, which was positioned outside the scanner. Two paradigms were used to evoke DOMS-associated muscle pain in the fMRI environment. The first paradigm involved alternate repeated active contractions of the right (sore) and the left (sound) quadriceps muscle group and was phasic in nature. The stimulus is identical to an isometric contraction and the evoked pain proves to be suited for the requirements of a functional MRI stimulus design that calls for stimuli with fast on- and offset. The whole paradigm consisted of 5 cycles of alternating non-painful (control condition) and painful contractions (stimulus) with eleven interleaved resting conditions (baselines), respectively 24 and 18 seconds Fig. 13; TR/TE  = 1950 ms/4.38 ms, FOV  = 256 mm, 256×256 matrix, 160 slices, FOV  = 256 mm, slice thickness  = 1 mm). Functional scans were acquired using a blood-oxygen-level-dependent (BOLD) protocol with a T2-weighted gradient echo-planar imaging (EPI) sequence . 20 axial slices were placed such that the best possible whole brain coverage was obtained, usually from the top of the cortex to the base of the cerebellum. The first three images were discarded to account for spin saturation effects thus eliminating non-equilibrium effects of the magnetization. MRI sequences were measured in the following order: anatomical scout, MPRAGE, and two EPI sequences. The first EPI was short and applied to accustom the subject to the noise. The second EPI consisted of 150 whole brain acquisitions divided into 5 cycles and entailing the protocol as described above. Eight sequences were acquired during the stimulus period time (24 s) and six during the interposed baseline periods  using a standard quadrature head coil. Preceding the functional measurements, a high resolution data set of each subject’s brain was acquired using a T1-weighted, 3D gradient-echo pulse sequence  are marked in 3. We focussed on clusters located in the following brain areas: frontal and parietal lobe, insula, cingulum, thalamus, basal ganglia and the cerebellum. Processing of the clusters included identification of Brodman's Area , determination of spatial centre (coordinates), and their statistical significance . Psychophysical data were recorded with custom made software.Imaging data analysis, registration, visualisation and statistical analyses were performed with the BrainVoyager QX software package . Pre-processing of the data was performed as previously described To quantify the pain, all subjects had to rate the pain perceived during contraction and physical stimulation of the sore quadriceps before the fMRI measurement. For this purpose the subjects were instructed to contract the painful muscles in 5 repeated cycles of 24 seconds each . All figures display significant focal activation with at least t>4.5.As expected, the vast majority of regions were activated by painful contraction or physical stimulation of the right quadriceps muscle group, while non-painful contraction or physical stimulation of the left thigh led primarily to activation in the respective somatotopically related areas of the primary sensory and motor area of the contralateral (right) cortex , 4. . The activation appeared as one large cluster not confined to the respective somatotopic area of the thigh but covering also the areas which are thought to correspond to the trunk and the lower limb. This is illustrated in , which shows a three-dimensional reconstruction of the activation pattern during DOMS-related pain projected onto the cortical surface for both means of stimulation, contraction  and physical stimulation . The more rostral located frontal lobe activation corresponds to the medial frontal gyrus which is the projection area of the supplementary motor area  and somatotopically related to the right lower extremity. Furthermore the activation expanded over the cingulate sulcus and merged with activation in the frontal lobe, the latter ones spanning the medial aspect of the fronto-parietal junction, i.e. the paracentral lobule, up to the medial frontal gyrus , 4.  SMA, , ., , . Surprisingly, the cortical surface area of S1 activated by painful contraction or painful physical stimulation was comparable to the area activated by non-painful physical stimulation . This was the contrary for M1, where non-painful means of stimulation only activated a very small area .In the left hemispheres the described clusters terminated for both, contraction and physical stimulation, more posterior in the medial parietal lobe with the centre located in the anterior precuneus, which represents the portion of the parietal lobe located caudal to the marginal sulcus and rostral to the parietooccipital sulcus , 4.We also found an increased pain-related BOLD-signal in the inferior parietal lobule for both contraction and physical stimulation IPL, see Table1,; t-values for activation in the anterior insula remained <5). Furthermore, in contrast to painful contraction, painful physical stimulation led to a large cluster of activation in the right-sided antero-medial insula.In the insular cortex, we found extensive left-sided activation with painful physical stimulation spanning from the anterior to the posterior part, whereas activation by painful contraction covered a much smaller area limited to the mid- and posterior insular cortex . A second cluster of activation was located in the left-sided mid-cingulate cortex , which was activated more extensively by physical stimulation than by contraction , . A se.Subcortical activations were confined to the thalamus, basal ganglia and the cerebellum see Table1..Activation in the cerebellum was confined to the right brain, ipsilateral to the painful stimulation. Remarkably, in the cerebellum painful contraction produced much larger clusters of activation, also expanding to the posterior lobe, while physical stimulation resulted only in a small cluster located in the anterior lobe In the present study, we describe widespread DOMS-related BOLD-signal increase in S1, M1 and the adjacent areas of the SMA, an area involved in the planning and execution of voluntary movements.The primary and secondary somatosensory cortex  are amongst those brain regions that consistently respond to painful stimulation with an increase in BOLD signal. In contrast to cutaneous pain, muscle pain leads to signal intensity increase in both the primary sensory and motor cortex (M1) as demonstrated previously using intramuscular electrical stimulation (IMES) and hypertonic saline Based on Penfield's somatotopic organization of S1 and M1 we expected activation primarily in the region around the vertex, which comprises those portions of the precentral and postcentral gyri that process sensorimotor signals related to the thigh. This was the case for painless contraction. Painful contraction and painful physical stimulation lead to more widespread activation of M1 and S1, extending to areas which are normally activated during sensory and motor stimulation of the upper and lower limb and the trunk. This widespread activation in presence of DOMS-related pain could underlie a transient reorganization of the somatosensory and motor cortex and be directly related to the painful experience, because activations in these areas are even larger during the more painful physical stimulation.DOMS is a subacute condition and usually entails pain for 24–72 hours that betimes results in maintenance of a protective posture. Previous studies dealing with chronic pain, such as the complex regional pain syndrome (CRPS), phantom limb pain and musculoskeletal pain conditions, like back pain, describe alterations in the somatotopic organization of the primary somatosensory cortex (S1). The amount of this reorganization was correlated with the subjective pain rating and these changes, which are based on processes of neuronal plasticity, could partially be reversed by analgesic interventions in back pain or progressive healing in CRPS Apart from that, we found activation in the precuneus, an area which belongs to the parietal association cortex. Here, activation may result from the inherent task, which required the recognition of the respective light signal (green or red) and its implementation into the appropriate motor action. Similarly, during physical stimulation the subjects could see the red and green light and anticipate the sequence of physical stimulation induced by the experimenter.Many previous pain studies (using both PET and fMRI) identified increases in regional cerebral blood flow (rCBF) or BOLD-signal in the cingulate cortex While the ACC is implicated in the processing of affective and reactive components of pain The cingulate motor area (CMA), forming part of the MCC, has previously been implicated in the processing of sensory stimulation, including pain of muscular origin The bilateral activation seen in the PCC may be related to pain, which is implied by its activation in response to both stimulation paradigms, but may also relate to the effort of intentionally inducing pain by active muscle contraction. The PCC maintains extensive parietal lobe connections and is thought to be involved in visuospatial orientation and assessment of self-relevant sensation In our study, DOMS-related pain evoked by self-controlled contraction led to large activation in the mid-posterior and posterior insula of the left brain, i.e. contralateral to the painful stimulus and a small area of activation in the right-sided anterior insula. In contrast DOMS pain evoked by physical stimulation led to activation overlapping with these areas including large clusters of activation bilaterally in the anterior insula. This is in agreement with previously reported findings that demonstrate the mid-posterior insula as constantly activated during tonic muscle pain Classically the STG  represents the primary auditory receptive cortex and auditory association areas. Further research indicates that the adjacent sulcus , the superior temporal sulcus (STS), is activated by the perception or observation of biologically relevant movements or by the perception of implied movements The inferior parietal lobule is an area that represents a boundary between secondary somatosensory areas (S2) and the parietal association cortex and is an area of sensory processing and sensorimotor integration Previous studies related to voluntary movements showed that unilateral movements result in a bilateral activation of M1 in the respective homotopic representations arguing for a principle of distributed processing in M1 According to previous findings, the ventral posterior complex in the thalamus is undoubtedly relevant for pain and temperature transmission in humans Concerning the cerebellum, pain-specific event-related imaging studies showed that certain cerebellar areas  seem to be involved in the processing of pain and the nociceptive leg withdrawal reflex Using event-related fMRI, the temporal evolution of brain activity during the subacute pain state DOMS was mapped in the human brain. We located strongest and widespread pain-related activations in the primary motor and sensory cortex that affected the area somatotopically related to the thigh and also adjacent areas reminiscent of the transient cortical remodelling described in chronic pain states, such as CRPS or phantom limb pain. Further pain-related activation was located in the SMA, IPL, STG, bilaterally in the insula and the cingulate cortex. Activation in the cerebellum was most widespread when pain from DOMS occurred in combination with limb movement. Our study demonstrated that defined stimulation or repeated contraction of a DOMS-affected painful muscle can evoke strong and reproducible increases in BOLD signal; therefore pain from DOMS provides an effective, non-invasive model to study the central processing of inflammatory muscle pain.


The practice of hemodialysis is a complex and technical high risk of infection to both the patient and the caregiver. Indeed, the infection is a major cause of morbidity and mortality in renal dialysis (15% of deaths).To evaluate the practices of hemodialysis center staff CHU Cocody.This is a cross-sectional study during the month of February 2012 with the health personnel in the hemodialysis center. Mainly nurses were interviewed. This study focused on the observation of indications for hand hygiene, compliance with asepsis, environmental control and risk prevention methods used.Almost all (96%) of the staff interviewed remove the jewelry on the hands and wrists before work. The equipment needed for dialysis is placed on a trolley that is regularly disinfected in 65% of cases. With regard to hand hygiene, hydroalcoholic solution is not available. After simple washing of hands, the staff uses alcohol for disinfection (55%) before wearing gloves. However, before going to another patient, staff dons new gloves. Disinfection is made one time (100%) and not four times. The arms and hands of patient with FAV are not washed before the meeting. Only the site is disinfected with alcohol before injection of the anticoagulant FDI. When connecting and disconnecting the generator, the gloves are also removed before further handling (83%). The screen is cleaned without being disinfected. The means of protection available at the Cocody Hemodialysis Center are essentially non-sterile gloves (80%) and sterile gloves (47%). Availability of masks is rare (5%); gowns and safety glasses are non-existent.This study identified the risk of infection in the Cocody hemodialysis center. The priority is mainly the provision of equipment necessary for patient safety.None declared


Current knowledge of saccade-blink interactions suggests that blinks have paradoxical effects on saccade generation. Blinks suppress saccade generation by attenuating the oculomotor drive command in structures like the superior colliculus (SC), but they also disinhibit the saccadic system by removing the potent inhibition of pontine omnipause neurons (OPNs). To better characterize these effects, we evoked the trigeminal blink reflex by delivering an air puff to one eye as saccades were evoked by sub-optimal stimulation of the SC. For every stimulation site, the peak and average velocities of stimulation with blink movements (SwBMs) were lower than stimulation-only saccades (SoMs), supporting the notion that the oculomotor drive is weakened in the presence of a blink. In contrast, the duration of the SwBMs was longer, consistent with the hypothesis that the blink-induced inhibition of the OPNs could prolong the window of time available for oculomotor commands to drive an eye movement. The amplitude of the SwBM could also be larger than the SoM amplitude obtained from the same site, particularly for cases in which blink-associated eye movements exhibited the slowest kinematics. The results are interpreted in terms of neural signatures of saccade-blink interactions. The neural mechanisms underlying the brainstem control of saccades have been well documented . BrieflyThe trigeminal blink reflex is the rapid and transient closure of the eyelids that is invoked most commonly by delivering an air-puff to one eye. It has thus far served as an under-appreciated but useful perturbation tool to test principles of the neural control of saccades. This manipulation has led to three interesting but seemingly incompatible results. One, a blink generated during fixation completely cease the tonic firing rate of OPNs The objective of the current study was to build on our knowledge of interactions between saccades and blinks. Our approach was to attempt to induce the trigeminal blink reflex during saccades evoked by sub-optimal microstimulation of the SC and to compare the metrics and kinematics of the stimulation-evoked eye movements with and without blinks. We hypothesized that SC inhibition of the blink system will not be potent when using sub-optimal microstimulation and therefore predicted that air-puffs, which are ineffective at producing blinks during supra-threshold SC microstimulation Macaca mulatta) underwent one or more surgeries in a sterile environment and under isoflurane anesthesia. The initial procedure consisted of placing a Teflon-coated stainless steel wire  under the conjunctiva of one eye and securing a head-restraint post to the skull. In the second procedure, one cylinder was cemented over a craniotomy. The chamber was placed stereotactically on the skull, slanted posteriorly at an angle of 38° in the sagittal plane. This approach allowed access to both colliculi and permitted electrode penetrations normal to the SC surface. After each surgery, the monkey was returned to its home cage and allowed to fully recover. Post-operatively, antibiotics and analgesics were administered as indicated in the protocol. As a result of the surgically added eye coil and head chamber, all animals were housed individually for safety. The Division of Laboratory Animal Resources continually monitored the animals and provided enrichment in the form of toys, radio, and television. Feeding of nutrition enriched biscuits occurred twice a day as well as the administration of daily fruit, vegetables, and a foraging mix of dried treats. Water was also given to each animal daily.All procedures were approved by the Institutional Animal Care and Use Committee at the University of Pittsburgh and complied with the guidelines of the Public Health Service policy on Humane Care and Use of Laboratory Animals. Four juvenile, male rhesus monkeys . The site-specific vector was determined with high or supra-threshold stimulation conditions . If necessary, the depth of the electrode was adjusted to obtain the shortest possible latency of the stimulation evoked saccade (20–40 ms). Next, low stimulation settings were obtained by selecting lower current intensities, frequencies, or both that reliably produced movements (>90% probability of evoking movement). This experimental manipulation also reduced the amplitude of the movements consistently (∼15% or more change in amplitude), as described by previous studies Stimulation-only trials (20%): As described above, microstimulation was delivered during the gap period and produced stimulation-only evoked movements (SoMs). Puff-only trials (10%): A puff of air was delivered to one eye to produce blinks during the gap period. These trials allowed us to characterize the small, loopy eye movement that accompanies the blink, which we refer to as a blink related eye movement (BREM). Eyelid movements were recorded using a small Teflon-coated stainless steel wire that was taped to the eyelid of the eye not implanted with the scleral coil. The eyelid coil signal, described in arbitrary units, was amplified in software to clearly identify eye closure as deflections in the vertical channel. Stimulation with blink trials (20%): Microstimulation and air-puff were combined to incorporate the effect of a blink (and prolonged OPN cessation) on a stimulation-evoked saccade, which we refer to as stimulation with blink-evoked movement (SwBM). The air-puff was delivered at random times before and during the stimulation. Control trials (50%): These were standard gap trials without stimulation or puff.The next phase of the experiment was to investigate the effects of blinks on saccades evoked by sub-optimal stimulation parameters. Four types of gap trials were randomly interleaved to address this goal. Each trial was digitized and stored on the computer’s hard disk for off-line analysis. We used a combination of in-house software and Matlab 7.10.0 (R2011a). Horizontal eye position and vertical eye and eyelid position along with onset and offset times of the stimulation train were stored with a resolution of 1 ms. Component velocities were obtained by differentiating the eye and eyelid position signals. Onset and offset of stimulation-evoked saccades and blink-related eye movements were then detected using a standard 30°/s velocity criteria, respectively.o to 40o in amplitude and −70o to 40o in direction. When paired with supra-threshold stimulation, air-puffs were completely ineffective in producing blinks for 16 of the 42 sites. For the remaining sites air-puffs yielded blinks on less than 10% of the trials. The result is consistent with previous studies that reported the inability to evoke blinks with supra-threshold collicular stimulation Effects of the trigeminal-blink reflex on saccades evoked by supra-threshold and sub-optimal microstimulation parameters were tested on 42 SC sites in four animals . The site-specific vectors, upon rotating into the right hemifield to pool data from left and right SC, spanned approximately 8Another result that can be extracted from the spatial trajectories plots  is that We wondered whether the significant hypermetria observed for monkeys 1 and 2 could have been simply due to a linear superposition of the BREM contribution. To test for this, we subtracted the maximum horizontal and vertical excursions of BREMs collected during puff-only trials (see Methods) from the endpoints of SwBMs. While this step naturally reduces the amplitudes of the SwBMs and shifts them closer to the amplitudes of the SoMs  attenuated in monkeys 3 & 4 compared to the other two animals? Previous work In an attempt to identify additional trends that could account for the distribution of radial amplitude differences between SoMs and SwBMs, within an animal and across animals, we also correlated the differences with numerous saccade features , but found no statistically significant trends.We have shown that, when paired with SC stimulation, the trigeminal blink reflex can be triggered reliably only when using sub-optimal stimulation parameters. The combined saccade-blink movement displayed a deviation in the spatial trajectory, attenuation of the eye velocity profile, and an increase in movement duration. In two of the four animals, the amplitude of the stimulation-evoked movement was consistently larger in the presence of a blink; the increase in amplitude was not due to an additive effect of the BREM. This spectrum of effects can be accounted for by complex combination of excitatory and inhibitory interactions at various nodes of the oculomotor circuit .Previous studies have demonstrated that the trigeminal blink reflex is rarely evoked during supra-threshold stimulation of the SC Previous studies based on visually-guided saccades have provided evidence that the blink reflex and the eye movement associated with it impose a suppressive effect on the saccadic system. Behaviorally, this is evident from the attenuation in the kinematics and temporal features of eye velocity waveforms . At the The results of our current study suggest that the suppressive effect of blinks on the saccadic system also applies to movements evoked by sub-optimal stimulation of the SC. We have argued recently Omnipause neurons (OPNs) located along the midline in the oculomotor paramedian pontine reticular formation are traditionally considered to gate saccadic eye movements Reflexive blinks evoked during visually-guided saccades can exhibit pronounced attenuation in the velocity waveforms Now we consider how this idea extrapolates to blinks combined with saccades evoked by SC microstimulation. It is known that the tonic activity of OPNs ceases during saccades evoked by SC stimulation The results of Saccade generation requires the brainstem BG to decode population activity emanating from various oculomotor structures, including the SC. The vector summation with saturation (VSS) model proposes that the cumulative sum of the collicular output drives the BG until saturation constrains the sum and terminates the movement


There are different strategies to reduce BLV prevalence. Eradication by culling of infected animals is not economically sustainable in highly infected regions such as Argentina, US or Japan. Segregation of BLV-infected cows is expensive due to duplication of facilities. Finally, several candidate vaccines based on recombinant viral proteins were unsuccessful to protect from challenge.Facing these problems, we propose a novel strategy based on the use of a live-attenuated BLV provirus. The rationale behind this strategy relies on the deletion of genes required to induce pathogenesis leaving those involved in infectivity, resulting in an attenuated mutant with impaired transmissibility.In a first set of experiments, we showed that the mutant is infectious and elicits an efficient immune response in sheep (n=3) and in cows (n=9). Lack of spread to uninfected sentinels further supports the safety of the vaccine. Based on these promising results, a validation program in herd (n=105) is ongoing to evaluate the capacity of the candidate vaccine to protect from wild-type BLV infection. The following experiments are carried out: quantification of the proviral loads, assessment of immune response efficiency , measure of viral expression in vivo (qRT-PCR) and ex vivo (expression of Tax and p24gag) and determination of provirus clonality during infection.This data will be instrumental for understanding the basic mechanisms undergoing during BLV infection and for elaborating a novel vaccine.


The purpose was to determine the frequency of recurrent/residual papillomas in women who had previously undergone vacuum-assisted excision biopsy (VAB) of benign papillomas without atypia and to identify factors that may be associated with recurrence.Women who had undergone VAB of papillomas and had subsequent breast imaging were identified from hospital records. Papilloma size, VAB device used and number of cores was recorded and subsequent imaging reviewed. Possible associations between the likelihood of recurrent/residual papilloma and patient age, lesion size and excised volume to papilloma volume ratio were analysed using Mann-Whitney U tests.Thirty-four women had subsequent imaging available. Median follow-up was 1,052 days. Twelve women (35%) had evidence of a mass at the excision site, with a median time to identification of recurrent/residual papilloma of 1,369 days. In three of the six cases that underwent ultrasound the recurrence was larger than the original lesion. Three recurrences were removed at open surgical biopsy and one by further VAB. None of the 34 women were diagnosed with atypia or malignancy during follow up. No significant association was found between the risk of recurrence and the age of the patient, the size of the initial lesion or the ratio of the volume of tissue removed to the lesion volume.Recurrent/residual papillomas are common but most do not require further intervention. No definite risk factors were identified. VAB remains the excision method of choice for most benign papillomas without atypia.


The accuracy of ligand-protein docking may be affected by the presence of water molecules on the surface of proteins. Water can form complex bridging networks and can play a critical role in dictating the binding mode of ligands. A recent analysis of high-resolution crystal structures of ligand-protein complexes revealed that 85% of the complexes had one or more water molecules bridging the interaction between ligand and protein. For predicting the binding modes and energies of protein-ligand interactions, molecular docking methods are commonly used. In order to obtain an accurate complex geometry and binding energy estimation, an appropriate method for calculating partial charges is essential. AutoDockTools software, widely used as interface for preparing input files, utilizes the either Gasteiger or Kollman partial charge calculation method for both protein and ligand charge calculations. However, it has already been reported that more accurate partial charge calculation and as a consequence, more accurate docking can be achieved by using quantum chemical methods. In common practice so far, the quantum chemical partial charges were applied to the ligands for docking calculations. The newly developed Mozyme function of MOPAC2009 allows fast partial charge calculation of proteins by quantum mechanical semi-empirical methods. Thus, in the current study, we use the semi-empirical quantum-mechanical partial charge calculations to investigate the interaction energies and polarization effects of the various components of the binding pocket on a set of Cathepsin B protein.The docking accuracy was computed by using the original AutoDock scoring function with the set of 19 protein ligand complexes using Gasteiger, AM1 and PM3 partial charge calculation methods. This helped us to compare the effect of the partial charge calculation method on docking accuracy. It was seen that the docking accuracy in regard to complex geometry significantly increased when partial charges of the ligands and proteins were calculated with the semi-empirical PM3 method. Our results demonstrate that (i) the energetic of the key water molecule are more favorable for the binding site in the Cathepsin B protein (ii) Water bridging and triangle formation were seen between the key amino acid residue and the ligand (iii) The internal energy is significant factor for the binding modes of various ligands. It was also observed a statistically significant overall increase in accuracy when water molecules are included during docking simulations. Out of the 19 complexes analyzed in the course of our study, the geometry of 17 complexes were accurately calculated using PM3 partial charges, while the use of Gasteiger charges resulted in only 8 accurate geometries.Our findings indicate that the inclusion of water molecules in ligand-protein docking results in significant increases in docking accuracy when the use of quantum chemical partial charge assignment on both ligand and proteins for predicting the docking simulations.


This mini-review highlights the progress that enables new insights into bacterial export and periplasmic protein organization, as well as new biotechnological applications combining the advantages of the periplasmic production and the Aequorea-based fluorescent reporter proteins.The bacterial periplasm is of special interest whenever cell factories are designed and engineered. Recombinantely produced proteins are targeted to the periplasmic space of Gram negative bacteria to take advantage of the authentic N-termini, disulfide bridge formation and easy accessibility for purification with less contaminating cellular proteins. The oxidizing environment of the periplasm promotes disulfide bridge formation - a prerequisite for proper folding of many proteins into their active conformation. In contrast, the most popular reporter protein in all of cell biology, Green Fluorescent Protein (GFP), remains inactive if translocated to the periplasmic space prior to folding. Here, the self-catalyzed chromophore maturation is blocked by formation of covalent oligomers via interchain disulfide bonds in the oxidizing environment. However, different protein engineering approaches addressing folding and stability of GFP resulted in improved proteins with enhanced folding properties. Recent studies describe GFP variants that are not only active if translocated in their folded form  Aequorea fluorescent proteins are the most widely used reporter proteins in all areas of biology. Due to their unique independence from cellular chaperones and non proteinogenic compounds (other than molecular oxygen) for chromophore maturation [in vivo applications.Since its discovery in 1962 [turation  they outturation  and othein silico modifications. Advanced engineering led to the availability of Aequorea-based fluorescent protein tools with surprising complexity like the multi colour bimolecular fluorescence complementation (BiFC)-system where split-FPs reconstitute to fluorescent proteins with altered spectroscopic properties [The long history of biotechnological exploration and engineering lead to a variety of available GFP variants with mutations affecting the spectral properties and the brightness by improving chromophore formation, folding or solubility as well as the discovery and engineering of related proteins from other organisms, e.g. the anthozoan mFruit-family proteins . Howeveroperties –14 or thoperties –17.The structure of correctly folded GFP consists of an internal fluorophore surrounded by a tight beta-barrel . MaturatSo far the folding properties of GFP variants have been exploited to report on the folding status of a fusion partner  or to stvia posttranslational Sec or cotranslational SRP mediated transport [Only very recently a super folder variant of GFP (sfGFP)  has beenransport , 21 – exIn this mini-review we summarize folding enhancement studies and their implications and applications with regard to bacterial export machineries.Waldo et. al. described a dependency of the fluorescence signal of protein fusions on the solubility and folding of proteins fused to the N-termini of GFPs [The broad application range of GFP is the result of early engineering approaches and identification of beneficial mutations. Especially the S65T mutation improved the spectroscopic characteristics and fluorescence quantum yield , while t of GFPs . From th of GFPs , 32 aros of GFPs , 36–39.Several years later, a selection process similar as the one applied in the folding reporter studies to enhance folding and solubility of target proteins fused to GFP was utilized to improve folding of GFP itself . Here, aet al [A completely different approach to select for GFP variants with improved folding characteristics was used by Fisher et al   system is a bacterial protein export pathway distinct from the general secretory pathway (Sec) with the remarkable characteristic to mediate transport of largely or completely folded proteins across the cytoplasmic membrane. The substrates of this pathway are therefore predominantly proteins that require the incorporation of cofactors in the cytoplasm, need assistance of cytoplasmic chaperones for folding prior to export or are proteins that fold too fast and stable for the Sec intrinsic stabilization of the unfolded polypeptide state . The Tatd ssTorA  of the tvia ssTorA  pathway is the most utilized export pathway in almost all bacteria . Presecrytoplasm  might alytoplasm , 24 indiSuper folder GFP (sfGFP), however, is able to fold and mature into its fluorescent conformation unaffected from the periplasmic environment , 21, as via the Tat pathway, can now be performed recruiting the native signal sequences and exportable sfGFP as demonstrated e.g. for EnvC and Pal [The discovery of the super folder GFP mutations and the folding robustness they confer to GFP allows maturation in the periplasm and enabled diverse new applications. Periplasmic protein localization studies using GFP, previously only possible by targeting the analyzed proteins  and Pal . Further and Pal  making t and Pal  a proper and Pal , 29, 50


Suprascapular nerve neuropathy constitutes an unusual cause of shoulder weakness, with the most common etiology being nerve compression from a ganglion cyst at the suprascapular or spinoglenoid notch. We present a puzzling case of a man with suprascapular nerve neuropathy that may have been associated with an appendectomy. The case was attributed to nerve injury as the most likely cause that may have occurred during improper post-operative patient mobilization.A 23-year-old Caucasian man presented to an orthopedic surgeon with a history of left shoulder weakness of several weeks' duration. The patient complained of pain and inability to lift minimal weight, such as a glass of water, following an appendectomy. His orthopedic clinical examination revealed obvious atrophy of the supraspinatus and infraspinatus muscles and 2 of 5 muscle strength scores on flexion resistance and external rotation resistance. Magnetic resonance imaging showed diffuse high signal intensity within the supraspinatus and infraspinatus muscles and early signs of minimal fatty infiltration consistent with denervation changes. No compression of the suprascapular nerve in the suprascapular or spinoglenoid notch was noted. Electromyographic studies showed active denervation effects in the supraspinatus muscle and more prominent in the left infraspinatus muscle. The findings were compatible with damage to the suprascapular nerve, especially the part supplying the infraspinatus muscle. On the basis of the patient's history, clinical examination, and imaging studies, the diagnosis was suspected to be associated with a possible traction injury of the suprascapular nerve that could have occurred during the patient's transfer from the operating table following an appendectomy.Our case report may provide important insight into patient transfer techniques used by hospital personnel, may elucidate the clinical significance of careful movement of patients following general anesthesia, and may have important implications for patient safety techniques, including those outlined in the World Health Organization Surgical Safety Checklist program. Suprascapular nerve neuropathy is a relatively uncommon cause of shoulder weakness and may be overlooked during physical examinations . The mosA 23-year-old Caucasian man presented to an orthopedic surgeon with a history of left shoulder weakness of about eight weeks' duration. He complained of pain and inability to lift even minimal weight, such as a glass of water, following an appendectomy. The patient had experienced severe acute pain in the left shoulder and scapula on the third day following his operation; however, he had not paid much attention to it, as the pain gradually subsided within the next two weeks, during which time he was hospitalized for ten days post-operatively for high fever. Following his discharge from the hospital, he noticed left shoulder weakness with an inability to lift even small weights; therefore, he visited an orthopedic surgeon.His orthopedic clinical examination revealed that he had obvious atrophy of the supraspinatus and infraspinatus muscles Figure , as wellRoutine MRI of the shoulder was done using a 3-Tesla scanner three months following the patient's the initial symptoms. A series of T2-weighted/turbo spin echo fat suppression oblique coronal, oblique sagittal, axial T2-weighted/turbo spin echo oblique sagittal, proton density fat suppression, and T2-weighted/gradient echo axial imaging slices through the left shoulder were obtained using a phased array dedicated shoulder coil. These MRI scans showed diffuse high signal intensity within the supraspinatus and infraspinatus muscles on the T2-weighted/turbo spin echo fat suppression images and early signs of minimal fatty infiltration consistent with denervation changes. In addition, no compression of the suprascapular nerve in the suprascapular or spinoglenoid notch and no denervation changes of the teres minor muscle or subscapularis muscle were seen. The MRI findings are presented in Figures To our knowledge, this is one of the first reported cases of suprascapular nerve neuropathy most likely attributable to nerve injury that may have occurred during improper post-operative patient mobilization. Although injuries of the nerve are relatively uncommon, the most frequent cause of neuropathy is associated with mass compression usually due to a ganglion cyst or other soft tissue tumors . Other cOur case, although it could be categorized as iatrogenic if attributed to improper post-operative patient mobilization, was not associated with surgical interventions in the anatomic region as such cases have previously been reported in the medical literature -8. InsteOur patient was healthy and fit and exercised on a regular basis, and his medical history was unremarkable. On the basis of his recent post-operative pain and weakness, clinical examination, MRI findings, and EMG, we assume that the patient had developed a suprascapular nerve neuropathy. Although the precipitating event was not witnessed, we speculate that the diagnosed nerve injury occurred during his post-operative transfer from the operating table following an appendectomy.On the basis of this information, we postulate that the mechanism of injury might be attributable to the muscle relaxation associated with general anesthesia that the patient was under during his appendectomy. Such muscle relaxation might have exacerbated the range of shoulder movement (hyperabduction) during his transfer from the operating table, leading to the combination of a traction injury and the so-called "sling effect."Based on our assumptions, our presently reported case highlights the importance of using proper patient transfer techniques by hospital personnel, as well as the clinical significance of careful patient mobilization following operations in which the patient has been under general anesthesia, and it may also have important implications regarding patient safety techniques, including the World Health Organization Surgical Safety Checklist program.Written informed consent was obtained from the patient for publication of this manuscript and any accompanying images. A copy of the written consent is available for review by the Editor-in-Chief of this journal.The authors declare that they have no competing interests.CPE compiled the data and obtained and interpreted the MRI studies. TK performed EMG and interpreted the imaging studies. LC performed the clinical examination of the patient. CPE and ESS were the main contributors to the writing of the manuscript. All authors read and approved the final manuscript.


The rheumatoid polyarthritis is the most frequent chronic polyarthritis. It affects essentially the woman between 40 and 60 years. Rheumatic subcutaneous nodules and tenosynovitis are usually associated with seropositive symptomatic rheumatoid polyarthritis. It is, however, rare that they constitute the essential clinical expression of the disease. In this case, it makes dispute another exceptional form of rheumatoid arthritis such as rheumatoid nodulosis. A 60-year-old woman was hospitalized for tumefaction of the dorsal face of the right hand evolving two months before. The clinical examination found subcutaneous nodules from which the exploration ended in rheumatoid nodules with tenosynovitis. The evolution after four years was favourable under corticosteroid therapy, methotrexate, and colchicine. The rheumatoid polyarthritis (RP) is the most frequent shape of the chronic polyarthritis, affecting joints provided with a synovial membrane. It is characterized by often bilateral and symmetric arthritis of several articular groups touching preferentially wrists and hands, evolving by pushes towards the deformation and the destruction of affected joints articulations. It gets essentially women between 40 and 60 years. The rheumatic subcutaneous nodules and the tenosynovitis are usually associated in seropositive RP. It is, however, rare that they constitute the essential clinical expression of the disease. We report a particular observation by its mode of revelation and its clinical evolution. 60-year-old woman with history of asthma under a long-term inhaled corticoids, consulted for tumefaction of the dorsal face of the right hand evolving for two months without triggering factor neither fever nor change of the general state. The interrogation found a notion of ancient arthralgia of large and small joints without arthritis and without any other systematic sign. The physical examination noted the presence of multiple mobile painful nodules on irregular surface of the dorsal face of the right hand as well as the front homolateral arm Figures , making The rheumatoid polyarthritis (RP) is a chronic disease of complex and multifactorial determinism, unpredictable evolution, and sometimes difficult positive and differential diagnosis. The ancient ACR criteria of RP contain four articular clinical criteria on seven and remain debatable . RheumatClassically, numerous arguments plead for an early diagnosis of RP. Effectively, osteocartilaginous destructions appear early in the evolution of the disease , and they are irreversible , 5.The rheumatoid factor is mostly absent during the first six months of evolution of RP. However, the presence of anti-CCP antibodies has a 50% of sensibility in a beginner RP, but especially a high specificity (97%) . It was A polyarthritis miming one PR can reveal other chronic inflammatory rheumatisms or even a neoplasm particularly a malignant lymphoma . At our The imaging by magnetic resonance can improve the diagnosis of tenosynovitis during RP . In our The extra-articular appearances of the rheumatoid polyarthritis must not be underestimated especially since they can constitute—even rarely—the essential clinical expression of the disease.


Quantum dots are nanometre-scale semiconductor crystals with unique optical properties that are advantageous for the development of novel chemical sensors and biosensors. The surface chemistry of luminescent quantum dots has encouraged the development of multiple probes based on linked recognition molecules such as peptides, nucleic acids or small-molecule ligands. This review overviews the design of sensitive and selective nanoprobes, ranging from the type of target molecules to the optical transduction scheme. Representative examples of quantum dot-based optical sensors from this fast-moving field have been selected and are discussed towards the most promising directions for future research. QDs typically have very broad continuous absorption spectra, at wavelengths extending from the ultraviolet to the visible, depending on the particle size. The broad excitation and narrow size-tunable emission spectra , negligible photobleaching, and high photochemical stability are some of their remarkable properties ,2 Figur. FluoresQDs have bandgap energies that vary as a function of size . The bandgap energy corresponds to the minimum energy that must be provided to move an electron from the valence to the conduction band creating a hole behind. The created electron–hole pair, an exciton, may recombine immediately to produce heat or light with energy equal to the bandgap energy, but it is more likely that trap states within the material trap either the electron or the hole  Figure . TherefoDue to the availability of precursors and the simplicity of crystallization, Cd-chalcogenide nanocrystals have been the most studied colloidal QDs. Several synthetic methods have been reported since the first synthesis of monodisperse CdE  nanocrystals  and extein vivo labelling, imaging and therapeutic applications in biomedicine are in expansion arene that hosted acetylcholine molecules resulting in the selective quenching of QDs fluorescence [The fluorescence quenching of QDs has been used to detect a number of other compounds of biological interest, such as the diuretic drug spironolactone . In one rescence .QD-based optical sensors have been designed to monitor carbohydrates like maltose or glucose. Engineered MBP with an oligohistidine tail, labelled with an acceptor dye at the allosteric site , was noncovalently self-assembled on DHLA-capped QDs and applied in FRET-based assays for monitoring maltose . The ass2O2 produced during GOx-catalyzed oxidation of glucose. An electron-transfer reaction occurred at the surface of the CdTe QDs where H2O2 was reduced to O2, which in turn lied in electron/hole traps on CdTe QDs and could be used as a good acceptor, thus quenching CdTe QDs fluorescence. The produced O2 participated then in the catalyzed reaction of GOx, forming a cyclic electron-transfer mechanism of glucose oxidation, which was favourable for the whole reaction system [One of the many QD-based strategies developed to detect glucose was based on the covalent conjugation of glucose oxidase (GOx) to CdTe QDs. The mechanism for glucose sensing relied on the fluorescence quenching of CdTe QDs, which was caused by Hn system .7.QDs have extraordinary electronic and optical properties that have been investigated for over a decade, making them one of the most prolific nanomaterials. These semiconductor nanocrystals have a broad variety of applications that include their use as a new type of fluorescent probes as well as active components in nanostructure-biomolecule complexes. Various optical transduction schemes have already been successfully explored for sensing a wide range of molecules, allowing low levels of detection and reduced interference of other compounds in complex samples. Currently a reality, multiplex detection is an ever-expanding research tool for QDs in optical arrays. Nonetheless, the vast majority of applications occur in bulk solutions limiting the development of reusable sensors, and routine analytical applications are hindered so far due to their known high toxicity. These will certainly be two of the challenges tackled in the near future concomitant with the continuous progress in the synthesis and conjugation methods for increased stability, sensitivity and binding specificity of QD-nanoassemblies incorporated in chemical sensors and biosensors.


Breast cancer is common in women and its metastases involve the skin in approximately one quarter of patients. Accordingly, metastatic breast cancer shown to be cutaneous through histology must be distinguished from a wide variety of other neoplasms as well as the diverse morphologic variants of breast cancer itself.We report the case of a 61-year-old Caucasian woman with cutaneous metastases of a bilateral ductal breast carcinoma that in histopathological examination mimicked an adnexal neoplasm with sebaceous differentiation.Against the background of metastatic breast carcinoma, dermatopathological considerations of sebaceous differentiation of skin lesions are presented and discussed focusing on the rare differential diagnosis of sebaceous carcinoma of the breast. Skin metastases of malignant tumors arise principally when the diagnosis of the primary cancer has been previously established, and cutaneous metastases from internal malignancies are an infrequent, although not totally rare, phenomenon . In contCutaneous metastases of carcinomas are encountered in 0.7-9.0% of all patients with cancer in general . In the In 2010 Fernandez-Flores investigated 78 cutaneous biopsies from 69 patients and identified six histological patterns of cutaneous metastasis: nodular, diffuse, infiltrative, intravascular, bottom heavy, and top heavy [In breast carcinoma in particular there is a wide range of clinical presentation of skin metastases. Most metastases are observed on the chest wall; less common sites include scalp, neck, upper extremities, abdomen and back . In geneTracking the differentiation from primary cutaneous malignancies can be challenging due to the ability of the tumor cells to mimic specific dermal structures. Although most skin metastases show morphologic and immunohistologic features of the primary malignancy, they can also mimic other dermatological patterns on histology.The initial dermatologic consultation of our 61-year-old Caucasian female patient occurred two years ago when she presented with a reddish, indolent nodule of the scalp 5 mm in diameter with local alopecia that she had noticed for the first time four months before. A small punch biopsy of her scalp exhibited solid proliferations of monomorphous tumor cells with a cytoplasm rich in vacuoles and sebaceous differentiation. Subepidermal spreading of the cells was knobby; a sclerodermiform-like spreading was predominant within the reticular dermis. The cells expressed pancytokeratin (MNF116) and epithelial membrane antigen (EMA) but staining for BerEP4 and carcino embryonal antigen (CEA) was negative. Therefore, we initially established the diagnosis of a primary cutaneous carcinoma with sebaceous differentiation. Upon thorough review of our patient's personal history she informed us of a previous diagnosis of a poorly differentiated invasive solid ductal breast carcinoma of her left breast five years previously, which was positive for estrogen receptor (ER) and progesterone receptor (PR), but negative for human epidermal growth factor receptor 2 (HER-2/neu) (Figure Further examination of our patient was then initiated. It showed a second moderately differentiated invasive ductal breast carcinoma of her right breast with a sonographic tumor thickness of 5 mm Figure , left. CThe tumor of the scalp was surgically removed in our department. Histopathological examination of this tissue showed a solid tumor consisting of large monomorphous cell proliferations with sebaceous differentiation, similar to the features found in the previous biopsy Figure . The immOur patient received axillary right sentinel node biopsy, ablatio mammae right, and one cycle of chemotherapy with paclitaxel and bevacizumab, but died due to sepsis two months after the diagnosis of cutaneous metastatic breast carcinoma. Detailed clinical data are given in table We were able to establish the final diagnosis of metastatic breast cancer with the histologic appearance of a sebaceous differentiated primary cutaneous carcinoma. Our patient had bilateral ductal breast cancer with identical hormone receptor status within five years. It remains unclear whether the cutaneous metastasis originated from the initially diagnosed breast cancer of her left mammary or from the second ductal carcinoma of her right breast.Cutaneous metastatic breast cancer must be distinguished from a wide variety of other neoplasms using histology. In the case presented, the tumor cells imitated the histological and immunohistological pattern of a sebaceous gland neoplasm.Interestingly, sebaceous differentiation can also occur in variable morphologic types of breast carcinoma, such as infiltrating or invasive ductal carcinoma, adenoid cystic carcinoma as well as others . It was The first description of a mammary sebaceous carcinoma was made in 1977 as a variant of lipid-rich carcinoma of the mammary gland . The immRegardless of this histogenetic discussion, dermatopathologists must be aware of this opportunity for misdiagnosis of diverse sebaceous neoplasms of the skin. Histological mimicry can hamper the correct diagnosis in small biopsy specimens because the lesions cannot be evaluated as whole, dimensional structures. Therefore, suspicious lesions should be excised completely. Reactivity for adipophilin is of great advantage in this setting . AdipophClinically, our patient presented with a reddish nodule on her scalp that caused focal alopecia, which was misdiagnosed in the first biopsy specimen as primary carcinoma of the skin with sebaceous differentiation. Due to the uncommon differentiation of the cells and the sebaceous-like pattern, diagnosis of cutaneous metastasis of a breast carcinoma was hard to establish on the biopsy. Only after complete removal of the lesion and with knowledge of the whole history of our patient could we finally establish the diagnosis of metastatic breast cancer.The authors declare that they have no competing interests.Written informed consent for publication from the patient's next of kin could not be obtained despite all reasonable attempts. The case is important to public health and every effort has been made to protect the identity of our patient. There is no reason to believe that our patient would object to publication.CSLM did all the histological reports, performed the histological examination and was a major contributor in writing the manuscript. RK collected the patient's data and wrote parts of the manuscript. ZFT and EFS cared for the patient in the gynecology department and provided the images of the breast. TV approved the final manuscript. CP cared for the patient in the dermatology outpatient unit. All authors have read and approved the final manuscript.


Cavitation-mediated therapies for the brain, such as ultrasound-induced Blood-Brain barrier opening and sonothrombolysis for the treatment of stroke, are being increasingly investigated. Robust methods for monitoring and controlling cavitation are necessary for safe translation of these techniques into clinical practice. This talk will review our work detecting and mapping cavitation activity in the brain, as well as using the cavitation signals to control treatments. The potential application of these techniques for mapping the vasculature will also be discussed.


Carcinosarcoma (CS) is a rare mixed mesodermal malignancy most commonly affecting the female reproductive organs, respiratory tract, head, and neck. Though infrequent, it may affect the gastrointestinal tract, most often the oesophagus and only very rarely the rectum. Histologically, it is composed of two distinct elements of epithelial and mesenchymal origin. Clinically, it is a very aggressive tumour with many patients presenting with metastatic lymph nodes or distant metastases at the time of diagnosis. Prognosis is poor despite intervention with the majority of patients dying within six months. Due to the rarity of this condition, there are no specific treatment guidelines presently available. We describe the case of an 80-year-old patient with carcinosarcoma of the rectum with discussion of the immunohistochemistry and review the available literature pertaining to this rare presentation. An 80-year-old female presented with a four-week history of fresh rectal bleeding, associated with poor appetite and weight loss. There were no other symptoms relating to change in bowel habit. Past medical history included ischemic heart disease, atrial fibrillation, hypertension, Parkinson's disease, and hysterectomy several years previously for benign disease. Abdominal examination was unremarkable; however, a low rectal tumour was palpated anteriorly on digital rectal examination. Subsequent colonoscopy showed a large solitary polypoidal growth in the lower rectum. Biopsy of the lesion provided a histological diagnosis of CS based on the presence of two clearly identifiable components on HPE. The tumour consisted of malignant glands set within an atypical stroma consisting of fascicles of spindle cells with thin, oval tapered nuclei and eosinophilic cytoplasm with perinuclear vacuolation. The stromal spindle cells showed florid mitotic activity. Further immunostaining confirmed the nature of the two components with the glandular tissue staining positively for CK20 and CEA. The stromal component did not stain positively for epithelial markers and was negative for cytokeratins. Staging computed tomography (CT) scan of the chest, abdomen, and pelvis and pelvic magnetic resonance imaging (MRI) were performed . A low rFollowing a five-week course of preoperative radiotherapy, the patient underwent radical laparoscopic-assisted abdominoperineal excision of the rectum (APER) with curative intent. Postoperative recovery was uneventful. Histological examination of the resected tumour revealed small amount of residual disease consisting largely of a poorly differentiated adenocarcinoma with a cellular stroma. The malignant tumour showed distinct carcinomatous  and sarcGiven the heterogeneous nature of the tumour, the two components showed distinct immunohistochemistry. The epithelial component stained positively for CK20  and CEA Eight-month follow-up did not show any evidence of recurrence and she was clinically well; however, she subsequently developed locoregional recurrence with pulmonary metastases at 16 months. In view of her frailty and significant cardiac risk factors, she was not considered for chemotherapy and succumbed to the disease 25 months after surgery.The term carcinosarcoma is used to describe a rare mixed mesodermal malignancy consisting of an epithelial component most often a mid to high grade adenocarcinoma and a sarcomatous element of mesenchymal origin which may be differentiated or undifferentiated . CS dispThe presentation and malignant behavior of CS is very similar to that of adenocarcinoma with the carcinomatous component showing a strong tendency to metastasise to lymph nodes and distant sites whereas the sarcoma plays only a very minor role in metastatic spread . PatientThe histogenesis of carcinosarcoma is poorly understood and multiple hypotheses have been proposed. Further characterization of the nature of the tumour itself has been achieved through immunohistochemistry, confirming the presence of distinct components. The most commonly observed pattern of staining is reactivity in the adenocarcinomatous component to the epithelial markers, CK20 and CEA , 9. The Within the large bowel, there are reports of carcinosarcoma from the caecum to the rectum; however, there does not appear to be any significant difference in terms of the potential to metastasize or the overall prognosis . A reviePrognosis of CS within the colon is poor with the longest reported survival being 49 months , 10–12 . Given tEvidence supporting the addition of adjuvant chemoradiotherapy in terms of improving long term outcome is lacking; however, the addition of RT for the patient in the current case did result in downstaging of the disease, allowing for resection with curative intent. Chemotherapy was not a viable option due to her significant cardiac comorbidities. As was done in this patient, strict follow-up is essential given the aggressive nature of this malignancy to detect any local recurrence or metastasis. Although this patient succumbed to the disease after 25 months, she remained disease-free until 16 months post-op which is significantly longer than all other reported cases of rectal carcinosarcoma.Rectal carcinosarcoma is a rare, aggressive mixed type of malignancy with mesenchymal and epithelial components. Immunohistochemistry is the gold standard for diagnosis, identifying two key features: differential staining of the distinct components and the lack of staining for epithelial markers in the sarcomatous component. Aggressive treatment similar to adenocarcinoma protocols with strict follow-up can improve survival rates.


Mucocele, a common benign cystic lesion of minor salivary gland and associated ducts develops following extravasation or retention of mucous material in the subepithelial tissue. Occurrence of mucocele of tongue is considered less frequent when compared to a higher incidence of mucocele in the lower lip of young patients. Different modalities of treatment, such as conventional surgical excision followed by newer techniques like cryosurgery, electrocautery have been proposed to completely remove the lesion and reduce the chances of recurrence. Herewith, we report a novel treatment technique using alginate impression material to aid in complete excision of mucocele of glands of Blandin-Nuhn.How to cite this article: Kumaresan R, Karthikeyan P, Mohammed F, Fairozekhan TA. A Novel Technique for the Management of Blandin-Nuhn Mucocele: A Case Report. Int J Clin Pediatr Dent 2013;6(3):201-204. They commonly appear as translucent bluish nodules that arise mostly on the lower labial mucosa followed by tongue, but other regions of oral cavity may also be affected.24The variation in color depends on size of lesion, its proximity to mucosal surface and elasticity.5Mucoceles of the glands of Blandin-Nuhn are exophytic and may resemble pyogenic granulomata, polyps or squamous papillomata. Clinically, a mucocele shows a positive history of trauma in most cases, rapid onset, alterations in size, bluish color and fluid-filled consistency.8Treatment usually consists of surgical excision of the lesion including the involved minor salivary gland. While other techniques, such as cryosurgery and laser ablation were indicated for complete removal of the lesion.The present report discusses a case of mucocele from glands of Blandin-Nuhn treated by surgical excision, where an unexpected surgical difficulty was overcome.A 9-year-old boy was brought to our Oral Surgery Clinic by his mother with the chief complaint of swelling on the ventral aspect of the tongue of 1 month duration. The swelling had gradually increased in size and interfered with speech and mastication. There was no history of trauma to the area, and the medical history was not significant.On clinical examination, a bluish, nonulcerated, oval shaped mass measuring about 25 × 15 mm in size was noticed on the left ventral aspect of the tongue. The swelling was painless, soft and mucosa over the lesion was tensed . On aspiSurgical removal of the lesion was planned under local anesthesia with 2% lignocaine. A vertical incision was made on the ventrum of the tongue and excision of the lesion was attempted. In course of dissecting the lesion from the underlying tissue, the cystic lining ruptured leading to an immediate extravasation of mucos . This main toto together with the associated minor salivary glands  by definition, is a cavity filled with mucos. They commonly present as either the mucous extravasation type or mucos retention type. The former is the primary cause of mucocele formation involving exudation of mucos into the connective tissue and surrounded by a granulation tissue envelope. The mucos retention type on the other hand appears due to a decrease or absence of glandular secretion produced by blockage of the salivary gland duct.1413Diagnosis is principally clinical. The appearance of mucocele is pathognomonic and the following data are considered helpful in the clinical diagnosis: a history of trauma, rapid onset, variation in size, a fluid-filled consistency, bluish color and recovery of mucos with aspiration.6 laser ablation7, micromarsupialization and steroid injection.8 Baurmash5 has advised on completely unroofing the lesion along its entire periphery to visualize and remove all of the glands present to prevent recurrence of the lesion.Small mucoceles are treated by surgical excision which includes the servicing mucous glands with evacuation of its contents. On the other hand large lesions may be treated by cryosurgery,2 In addition, incomplete excision of the cystic lining may also result in recurrence of the lesion. Hence, clearly demarcating the limit of the lesion is of prime importance in order to make the surgical excision complete and easier. Filling the cystic cavity with ultraflow rubber based impression material or alginate impression material presurgically improves the visual access for surgical excision.14 However, use of alginate is not recommended in areas where extension into surrounding tissue planes is possible leading to a foreign-body reaction in the tongue as a result of residual alginate within the tissue.15During surgical excision on the ventral surface of the tongue, all the minor salivary gland tissues in the surgical field may be electively excised to avoid iatrogenic recurrence of a mucocele by residual salivary gland tissue left behind.In our case, during surgical exploration of the mucocele an unexpected rupturing of the lesion had occurred making the surgical exploration difficult. A thin mix of alginate impression material injected into the lesion clearly delineated the entire lesion and complete excision was made possible. No foreign-body reaction was noted post-operatively.The use of alginate impression material to define the boundaries of the lesion is an effective, safe and economical way of treating an anterior lingual salivary gland mucocele, whilst ensuring complete removal and minimal chance of a recurrence. If performed correctly, this technique will undoubtedly prove to improve the surgical compliance thereby curtailing the recurrence of mucocele in the oral cavity.


AbstractPoaceae bioinformatics. GrassBase and GrassWorld can continue to grow productively if the proliferation of alternative classifications and datasets can be brought together into a consensus system. If the datasets are reconciled instead of diverging further apart a long term cumulative process can bring knowledge together for great future utility. This paper presents the Poaceae as the first and largest model system for e-taxonomy and the study of classification development in plants. The origin, development, and content of both datasets is described and key contributors are noted. The challenges of alternative classifications, data divergence, collaborative contribution mechanisms, and software are outlined.GrassBase and GrassWorld are the largest structured descriptive datasets in plants, publishing descriptions of 11,290 species in the DELTA format. Twenty nine years of data compilation and maintenance have created a dataset which now underpins much of the  Grasses and floristic knowledge. Grasses (Poaceae) are undisputedly the most economically important family of flowering plants (www.fao.org), including wheat (Triticumaestivum L.), rice (Oryzasativa L.), maize (Zeamays L.), sugar cane (Saccharumofficinarum L.), bamboos, forage grasses, and lawn grasses. As a group so fundamental to the human civilisation the grasses have always been well studied and the cumulative body of knowledge on the circa 12,000 species  a radical modernisation of web presentation will be necessary, including links to observation data, plant ontogenies, and provision of machine readable data output.Poaceae taxonomic community are needed to take grass e-taxonomy forwards. Failure to plan and collaborate could lead to an increasing proliferation of contradictory classifications. Unique datasets of great value could be rendered obsolete if software development and database maintenance does not keep pace with technology platforms. Investment in community database integration and infrastructure could unlock untapped research and data mining potential of many historic datasets. The rich data and the long history of database compilation in the grasses present an unprecedented opportunity to study the development of classifications and to develop e-taxonomic models. The authors would like to invite potential collaborators to discuss dataset improvements and future plans.This paper argues that a collaborative approach and careful thought across the PageBreak


The Coral Triangle in the Indo-Pacific is a region renowned for exceptional marine biodiversity. The area could have acted as a ‘centre of origin’ where speciation has been prolific or a ‘centre of survival’ by providing refuge during major environmental shifts such as sea-level changes. The region could also have acted as a ‘centre of accumulation’ for species with origins outside of the Coral Triangle, owing to it being at a central position between the Indian and Pacific oceans. Here, we investigated support for these hypotheses using population-level DNA sequence-based reconstructions of the range evolution of 45 species (314 populations) of Indo-Pacific reef-associated organisms. Our results show that populations undergoing the most ancient establishment were significantly more likely to be closer to the centre of the Coral Triangle than to peripheral locations. The data are consistent with the Coral Triangle being a net source of coral-reef biodiversity for the Indo-Pacific region, suggesting that the region has acted primarily as a centre of survival, a centre of origin or both. These results provide evidence of how a key location can influence the large-scale distributions of biodiversity over evolutionary timescales. Global-scale analyses of marine diversity show that some locations are unusually rich in species  and knowThere is evidence that the Coral Triangle region is a contact zone of taxa for some species that have diversified in allopatry , wherea2.http://www.ncbi.nlm.nih.gov/genbank/) for broadly distributed Indo-Pacific reef-associated fish and invertebrates with a partial distribution in the Coral Triangle region. We used data for species that had sequences from three or more locations, where eight or more individuals had been sampled from each, and where at least one of the locations sampled was within 2500 km of our approximated centre of the Coral Triangle . The slope of the relationship between geographical distance from the centre of the Coral Triangle and the relative time of population establishment was significantly negative . Populations undergoing the earliest establishments (oldest) were closer to the centre of the Coral Triangle than those undergoing the latest establishments (youngest) . Together these results support the hypothesis that the Coral Triangle has acted as a net source of extant Indo-Pacific coral-reef diversity, either by acting as a centre of origin and/or a centre of survival. Notably, time-calibrated phylogenies indicate that Indo-Pacific reef fish species tend to have diverged from their sister taxa on average approximately 3 Ma [There was a consistent pattern of the populations undergoing earlier establishment (oldest) being closest to the estimated centre of the Coral Triangle  and habitat specialization . DemograBuilding on previous studies, our results support the concept that refugia have a pivotal role in the recovery of communities following Pliocene and Pleistocene habitat loss. This emphasizes the importance of refugia for preventing biodiversity loss and has relevance to ongoing threats to shallow water reef communities through habitat destruction, ocean acidification and thermal stress linked to climate change . It has


Recurrent infections are very common in children and a major challenge for pediatricians. They affect the children’s quality of life, cause absences from school and lost parental working days, and repeated medical examinations, hospital admissions as well as antibiotic therapies lead to high costs for society. Given their prevalence and clinical importance, various prevention strategies have been developed.Some of these strategies have considered that innate and adaptive immunity are strictly related with gut microbiota and interact in the modulation of host’s reactivity to infections. Variations in gut microbiota related to age, nutrition or underlying disease may influence immune system defenses against viruses and bacteria. Furthermore, disturbed gut colonization patterns are proposed to be associated with the development of allergic disease. This explains why probiotics represent an important option in the prophylaxis and the management of recurrent infectious diseases as well as allergic diseases. However, the incomplete understanding of what constitutes a healthy gut microbiota that promotes tolerance, remains a challenge. Further understanding of gut microbial functions may pave the way for more effective prevention and treatment strategies.in vitro and in vivo. Pidotimod (3-L-pyroglutamil-L-thiaziolidine-4-carboxylic acid) is a synthetic dipeptide which stimulates the phagocytic activity of polymorphonuclear leukocytes, enhances the killing activity of human alveolar macrophages, increases the cytotoxic activity of natural killer cells, and stimulates cytokine production. It also influences the maturation and activity of dendritic cells by increasing the expression of the key surface markers that help to trigger T cell activation and increased IL-12 production. The majority of studies have shown that the number of infections decreases after pidotimod treatment, but they are affected by some methodological weaknesses. Further studies are urgently needed to confirm the pediatric population that should have the greatest benefit with the administration of immunostimulants.Another approach used for prevention of recurrent infections is represented by the administration of immunostimulants: i.e. molecules of bacterial or synthetic origin that interact with immunological mechanisms More studies are needed to identify the most promising probiotic strains and study populations, and to evaluate the mechanisms behind the possible effects of probiotics on OM.


The aim of the present study was to establish human ovarian stroma within the mouse subcutaneously, in order for the resulting stroma to serve as a useful preclinical tool to study the progression of human ovarian cancer in a humanized ovarian microenvironment. Normal human ovarian tissues were subcutaneously implanted into severe combined immunodeficient (SCID) mice and then the implants were identified by immunohistochemistry. The implants became vascularized and retained their original morphology for about 4 weeks following implantation. Immunohistochemical staining for cytokeratin-7 confirmed the ovarian origin of the epithelial cells. CD34 staining demonstrated human-derived vessels. Positive estrogen receptor and partially-positive progesterone receptor staining indicated the estrogen and progesterone dependence of the implants. Only vascular pericytes expressed α-smooth muscle actin, indicating the normal ovarian origin of the xenografts. Human ovarian tissue successfully survived in SCID mice and retained its original properties. This humanized mouse model may be used as preclinical tool to investigate ovarian cancer. The mice were housed under specific pathogen-free, temperature-controlled conditions. Their cages, bedding and drinking water were autoclaved and changed regularly. Food was sterilized by irradiation. The mice were maintained in a daily cycle of alternating 12-h periods of light and darkness. All experimental procedures were conducted according to the Guide for the Care and Use of Laboratory Animals and were approved by the Animal Care and Use Committee of Nanjing Medical University .Normal, human, noncancerous ovarian tissues were obtained from patients that had undergone a total hysterectomy with prophylactic oophorectomy at the First Affiliated Hospital of Nanjing Medical University. The use of human samples in the study was approved by the Committee for Ethical Review of Research at Nanjing Medical University, according to the ethical guidelines of the Declaration of Helsinki and each patient provided informed consent to participate in the study.Under sterile conditions, normal human ovarian tissues were sliced into small sections (~4×4×4 mm in size). Three small sections of normal human ovarian tissue were randomly selected for subsequent histological examination; the other pieces were stored in phosphate-buffered saline at 4°C until implantation into the mice.Prior to implantation, the mice were anesthetized with an intraperitoneal injection of 1% pentobarbital sodium . The surgical procedure was performed as previously described, with certain modifications ,10. In bFollowing implantation, the ovarian xenografts were subjected to weekly gross examinations. Mice were sacrificed at one, two or four weeks following surgery, and the human xenografts, including the mouse tissue surrounding the implanted human ovarian tissues, were harvested for histological assessment and immunohistochemical analysis. The harvested specimens were fixed in 10% formalin for examination.Specimens were dehydrated and embedded in paraffin. Sections (4-μm thick) were stained with hematoxylin and eosin and examined under a microscope. For immunohistochemical staining, the specimens were deparaffinized and rehydrated using xylene and graded alcohol. Sections were mounted on slides, subjected to antigen retrieval and then incubated with rabbit monoclonal antibodies against human estrogen receptor (ER), progesterone receptor (PR) and CD34 , as well as human cytokeratin (CK)-7, CK-20 and α-smooth muscle actin . Immunocomplexes were visualized using the diaminobenzidine method and sections were counterstained with hematoxylin. Negative controls were serial sections processed without the primary antibody. These reagents exhibited no cross-reactivity between the relevant species (mouse and human). All histological examinations were performed by one experienced pathologist.From one week following the implantation, the ovarian xenografts were subjected to weekly gross examinations. The embedded human ovarian tissues survived in the SCID mouse hosts for up to six weeks. No infection or tissue rejection reactions were observed at the time of tissue harvest, and all the implants became vascularized and survived well on the sheath of mouse muscles in the subcutaneous tissue .Histological evaluation revealed that the morphology of the human ovarian tissues embedded in the SCID mice was similar to the morphology of the original tissues prior to implantation. The viability of the cells within the xenografts was confirmed by the intact nature of the cell membranes, normal granulation of the cytoplasm and normal size and staining characteristics of the nucleus . In a nuImmunohistochemical analysis demonstrated that the harvested specimens were positive for ER and partially positive for PR, indicating that the inoculated ovarian tissues had remained estrogen- and progesterone-dependent, similar to the original tissues . Positivin vivo analysis, and a number of important research advances have been made using mice as a model for the study of various biological systems. However, mice are not the same as humans, and in vivo studies of human biology are severely limited by ethical and technical constraints. There is a growing need for animal models, which can be used to construct in vivo studies of human cells, tissues and organs without putting individuals at risk. Humanized mice, or mouse-human chimeras, have been developed to overcome these constraints and are now an important research tool for the in vivo study of human cells and tissues (in vivo study of human biological processes, which may otherwise not be possible. In the present study, a novel protocol was developed for the establishment of normal human ovarian stroma within the subcutaneous tissues of mice. It was hypothesized that the implanted stroma would accurately mimic the human ovarian microenvironment, and result in a humanized mouse model that may potentially serve as a useful preclinical tool for investigating the progression of human ovarian cancer.Complex biological processes often require  tissues . Humaniz tissues . The devA key factor in the success of this engraftment and the long-term survival of human xenografts is the use of SCID mice. Failure to develop mature T and B lymphocytes and the lack of an immune system makes SCID mice an ideal model for cell and tissue-transfer experiments. As a classical model, the subcutaneous mouse model has been widely used for a number of years. In the present study, normal, human ovarian tissues were subcutaneously implanted into SCID mice with the aim of developing a novel human ovarian tissue-derived mouse model.Numerous studies have used mice with xenografts implanted under the renal capsule . HoweverA normal, human tissue microenvironment contains three-dimensional intercellular interactions between the stroma cells, epithelial cells and cytokines. These are essential for maintaining epithelial polarity and modulating the growth inhibition of normal cells ,19. HoweIn the present study, the epithelial cells in the xenografts were shown to be derived from normal ovarian tissues, as these cells were positive for CK-7, which is diffusely distributed throughout the cytoplasm of ovarian cells. Furthermore, human-derived vessels within the implants remained viable for a period of four weeks following implantation. Positive staining for the ER and partially positive staining for the PR indicated that the inoculated ovarian tissues continued to be estrogen- and progesterone-dependent. α-SMA was found to be expressed only in the vascular pericytes and not in the epithelium or stromal fibroblasts, indicating that the subcutaneous xenografts of human ovarian tissues in the SCID mice consisted of cells derived from normal tissues. These observations indicated that the implanted human ovarian tissues survived well in the SCID mice.The results of the present study indicate that the humanized SCID mouse ovarian transplant model has the potential to serve as a useful preclinical tool, which may be used to build a humanized mouse model of ovarian cancer to investigate the efficacy of chemotherapeutic strategies. The absence of an immune system is one of the major limitations inherent in this type of model. However, SCID mice are likely to continue to be critical in establishing xenograft models that recapitulate the tumor growth and spread observed in patients.In conclusion, the present study demonstrated that human ovarian tissue successfully survives in a SCID mouse host and retains the properties of the original normal ovarian tissues. In addition, the experimental protocol used in the current study may be used to establish a humanized mouse model of ovarian cancer for preclinical research.


Case. This case report describes the operative management of 16-year-old male with a symptomatic chondroblastoma of the distal femur with breach of the chondral surface. Following appropriate imaging and core needle biopsy, the diagnosis was confirmed histologically. The patient then underwent intralesional curettage and osteochondral allograft reconstruction of the defect. At one-year follow-up the patient was pain-free and has obtained excellent range of motion. There is radiographic evidence of allograft incorporation and no evidence of local recurrence. Conclusion. Osteochondral allograft reconstruction is an effective option following marginal resection and curettage of chondroblastoma involving the chondral surface of the distal femur. Chondroblastoma was first described by Codman in 1928 as “epiphyseal chondromatous giant cell tumors of the proximal humerus” –6. SubchIn August of 2012 a 16-year-old male presented complaining of rest and activity pain worsening for the past year. He denied any history of trauma. The patient reported is worsening left knee pain and decreased range of motion were limiting his daily activities. Radiographs from an outside facility (not available for publication) showed a lobulated well-defined epiphyseal lesion with sclerotic margins and without stippled calcifications. Computerized tomography (CT) further defined a low-attenuation lesion in the medial femoral condyle measuring 13 mm by 16 mm with well-defined sclerotic margins and depression of the subchondral bone at the lesion border . DiffereThe patient underwent resection and allograft reconstruction in January of 2013. The posterior aspect of the medial femoral condyle was approached through a posterior medial approach to the knee using an extensile skin incision dissecting down through the interval between the medial head of the gastrocnemius and the semimembranosus tendon as described by Burks and Schaffer . The lesThe patient tolerated the procedure well without complication and was instructed to remain touch down weight bearing in a hinged knee brace locked in full extension for two weeks. He was seen two weeks postoperatively with minimal pain and swelling noted on exam. At this point the Bledsoe brace was unlocked and he was allowed to weight bear as tolerated. He then completed a full course of physical therapy and was seen in follow-up at the three- and five-month marks. At the five-month postoperative visit he denied any pain and reported ability to run and jump without pain or limitation. He demonstrated full active flexion and extension compared to the contralateral extremity . At his Chondroblastoma is a rare benign primary bone tumor commonly presenting in the knee of young patients , 5. GiveWe report a case of chondroblastoma involving the posterior surface of the medial femoral condyle in an otherwise healthy and active 18-year-old male. After obtaining appropriate imaging and a tissue diagnosis via core needle biopsy, marginal excision and osteochondral allograft reconstruction was performed in an attempt to prevent recurrence and maintain joint integrity. At the one-year mark there was no evidence of disease recurrence and the patient has achieved full painless, active range of motion. He has returned to school and sport and is very satisfied with the outcome of his treatment. At one year the osteochondral allograft has incorporated and remained stable and flush with the remaining articular surface, which should limit the progression of the degenerative process. At this point we will continue to follow up the patient for evidence of local recurrence, metastasis, and degenerative change with serial radiographs. We would expect our patient's risk of local recurrence and metastatic disease to be similar to rates reported in the literature as we treated the lesion with standard intralesional curettage and local adjuvant therapy. Our focus, as it relates to this case report, will be long-term follow-up of osteochondral allograft incorporation and progression of degenerative disease.Given our experience with this case we feel that intralesional curettage with osteochondral allograft reconstruction is a viable surgical option for chondroblastomas involving the articular surface of the knee. This reconstruction technique allows for painless weight bearing and full range of motion.