FOXP3 expression in cancer cells and anthracyclines efficacy in patients with primary breast cancer treated with adjuvant chemotherapy in the phase III UNICANCER-PACS 01 trial

BackgroundPredictive markers of response to chemotherapy are lacking in breast cancer patients. Forkhead Box Protein 3 (FOXP3) is an anti-oncogene whose absence in cancer cells could confer resistance to DNA damaging agent. So we made the hypothesis that FOXP3 expression predicts the response to anthracyclines in breast cancer patients and that adjuvant chemotherapy adding taxanes to anthracyclines confers an overall survival (OS) benefit over anthracyclines alone, in patients with FOXP3-negative tumors.Patients and methodsExpression of FOXP3 in cancer cells was evaluated by immunohistochemistry in tumor samples from 1097 patients who participated in the PACS01 randomized trial that evaluated in adjuvant setting the adjunction of docetaxel (Taxotere) to anthracyclines in patients with localized breast cancer. Kaplan–Meier analysis and Cox regression model were used to assess OS according to the presence or absence of FOXP3 expression in tumor cells.ResultsFour hundred and five tumors were found to express FOXP3 (37%). FOXP3 expression in breast cancer cells was associated with better OS (P = 0.003). Uni- and multivariate survival analyses according to treatment arm revealed that FOXP3 expression in breast cancer cells is independently associated with improved OS in patients treated with anthracycline-based adjuvant chemotherapy, but not in patients treated with sequential anthracycline–taxane. Moreover, in vitro experiments showed that FOXP3 induction in breast cancer cell lines using histone deacetylase inhibitor enhances anthracyclines efficacy.ConclusionFOXP3 expression in tumor cells may be an accurate predictive biomarker of anthracycline efficacy in breast cancer.     

Spinocerebellar ataxia type 4 (SCA4): Initial pathoanatomical study reveals widespread cerebellar and brainstem degeneration

Summary. Spinocerebellar ataxia type 4 (SCA4), also known as ‘hereditary ataxia with sensory neuropathy’, represents a very rare, progressive and untreatable form of an autosomal dominant inherited cerebellar ataxia (ADCA). Due to a lack of autopsy cases, no neuropathological or clinicopathological studies had yet been performed in SCA4. In the present study, the first available cerebellar and brainstem tissue of a clinically diagnosed and genetically-confirmed German SCA4 patient was pathoanatomically studied using serial thick sections. During this systematic postmortem investigation, along with an obvious demyelinization of cerebellar and brainstem fiber tracts we observed widespread cerebellar and brainstem neurodegeneration with marked neuronal loss in the substantia nigra and ventral tegmental area, central raphe and pontine nuclei, all auditory brainstem nuclei, in the abducens, principal trigeminal, spinal trigeminal, facial, superior vestibular, medial vestibular, interstitial vestibular, dorsal motor vagal, hypoglossal, and prepositus hypoglossal nuclei, as well as in the nucleus raphe interpositus, all dorsal column nuclei, and in the principal and medial subnuclei of the inferior olive. Severe neuronal loss was seen in the Purkinje cell layer of the cerebellum, in the cerebellar fastigial nucleus, in the red, trochlear, lateral vestibular, and lateral reticular nuclei, the reticulotegmental nucleus of the pons, and the nucleus of Roller. In addition, immunocytochemical analysis using the anti-polyglutamine antibody 1C2 failed to detect any polyglutamine-related immunoreactivity in the central nervous regions of this SCA4 patient studied. In view of the known functional role of affected nuclei and related fiber tracts, the present findings not only offer explanations for the well-known disease symptoms of SCA4 patients (i.e. ataxic symptoms, dysarthria and somatosensory deficits), but for the first time help to explain why diplopia, gaze-evoked nystagmus, auditory impairments and pathologically altered brainstem auditory evoked potentials, saccadic smooth pursuits, impaired somatosensory functions in the face, and dysphagia may occur during the course of SCA4. Finally, the results of our immunocytochemical studies support the concept that SCA4 is not a member of the CAG-repeat or polyglutamine diseases. Y. Hellenbroich and K. Gierga are joint first authors C. Zühlke and U. Rüb are joint senior authors     

Diffusion tensor MRI shows abnormal brainstem crossing fibers associated with ROBO3 mutations

Horizontal gaze palsy with progressive scoliosis (HGPPS) is caused by mutations in the ROBO3 gene, critical for the crossing of long ascending medial lemniscal and descending corticospinal tracts in the medulla. Diffusion tensor imaging in a patient with HGGPS revealed the absence of major pontine crossing fiber tracts and no decussation of the superior cerebellar peduncles. Mutations in the ROBO3 gene lead to a widespread lack of crossing fibers throughout the brainstem.     

Accurate placement of bone tunnels in reconstruction of the anterior cruciate ligament - a contribution of computer-assisted navigation

Rupture of the anterior cruciate ligament (ACL) is one of the most frequent injuries to the knee joint in the young. ACL repair is a major orthopedic procedure most often performed in the younger adult population. Early stabilization of the knee joint by ACL reconstruction also decreases the risk of injury to other important structures. At ACL reconstruction, the biggest problem is usually the exact placement of drilled tunnels. This significantly affects the outcome of surgery, i. e., range of motion, knee joint stability, reaction of the synovium in the knee, pain, impingement and potential graft failure with lesion development. However, 70 % of ACL reconstructions are carried out by orthopedic surgeons whose experience is limited to less than 20 ACL repair procedures in a year! Arthroscopy does not allow the surgeon to gain a complete 3D view of important anatomical structures, particularly in the anteroposterior direction. Computer-assisted navigation systems should aid in minimizing these problems. First reports on the use of computer-assisted navigation in ACL reconstruction, which have already been published in the international literature, have provided clear evidence that more exact bone tunnel placement can be achieved with navigation than with the use of conventional techniques. In addition, kinematic navigation enables us to measure anteroposterior and rotational knee stability, isometry, impingement and the angles of bone tunnel placement. It permits a choice from various types of graft. Last but not least, kinematic navigation provides a tool for recording surgery outcomes without a necessity to use further examination methods. Its drawbacks, namely, the learning curve, additional fixation of navigation probes to the femur and tibia and slightly longer operative time, should be considered in the context of presumed long-term benefits for the patient.     

家兔颈动脉粥样硬化狭窄简捷模型建立

目的探讨建立简捷、经济实用、稳定可靠的适于颈动脉粥样硬化性狭窄(CASS)的外科治疗研究的家兔CASS动物模型条件。方法新西兰白兔21只,采用3％过氧化氢液灌注动物的双侧颈总动脉,造成氧化应激损伤,然后以特定高脂饲料喂养动物不同时间。评价各组动物血管狭窄程度和病理改变特点。结果8周组和12周组中重度狭窄达到80％,个别血管出现闭塞。4周组轻度狭窄亦达到60％。病理检查证实高脂饲料喂养8周时,动脉的粥样硬化病理改变已属于纤维班快期。结论 按本实验方法,氧化应激损伤后喂养8周的动物颈动脉狭窄程度和病理改变程度已符合CASS外科治疗实验研究需要。     

半枝莲多糖的纯化及对S180荷瘤小鼠的抑瘤作用

目的将半枝莲粗多糖纯化后测定多糖含量,并比较粗多糖与精多糖的抑瘤率,为半枝莲化学及药理研究提供有益的参考。方法采用分光光度法、苯酚-硫酸显色法,在490nm处测定其含量;并利用体内实验测定多糖的抑瘤效果。结果精多糖的多糖含量为54．12％,平均加样回收率为99．66％,粗多糖的抑瘤率为45．96％,精多糖的抑瘤率为22．14％。结论测定多糖含量的方法操作简便,准确可靠,精多糖的抑瘤效果没有粗多糖抑瘤效果明显。     

Involvement of the auditory brainstem system in spinocerebellar ataxia type 2 (SCA2), type 3 (SCA3) and type 7 (SCA7)

Aims: The spinocerebellar ataxia type 2 (SCA2), type 3 (SCA3) and type 7 (SCA7) are clinically characterized by progressive and severe ataxic symptoms, dysarthria, dysphagia, oculomotor impairments, pyramidal and extrapyramidal manifestations and sensory deficits. Although recent clinical studies reported additional disease signs suggesting involvement of the brainstem auditory system, this has never been studied in detail in SCA2, SCA3 or SCA7. Methods: We performed a detailed pathoanatomical investigation of unconventionally thick tissue sections through the auditory brainstem nuclei (that is, nucleus of the inferior colliculus, nuclei of the lateral lemniscus, superior olive, cochlear nuclei) and auditory brainstem fibre tracts (that is, lateral lemniscus, trapezoid body, dorsal acoustic stria, cochlear portion of the vestibulocochlear nerve) of clinically diagnosed and genetically confirmed SCA2, SCA3 and SCA7 patients. Results: Examination of unconventionally thick serial brainstem sections stained for lipofuscin pigment and Nissl material revealed a consistent and widespread involvement of the auditory brainstem nuclei in the SCA2, SCA3 and SCA7 patients studied. Serial brainstem tissue sections stained for myelin showed loss of myelinated fibres in two of the auditory brainstem fibre tracts (that is, lateral lemniscus, trapezoid body) in a subset of patients. Conclusions: The involvement of the auditory brainstem system offers plausible explanations for the auditory impairments detected in some of our and other SCA2, SCA3 and SCA7 patients upon bedside examination or neurophysiological investigation. However, further clinical studies are required to resolve the striking discrepancy between the consistent involvement of the brainstem auditory system observed in this study and the comparatively low frequency of reported auditory impairments in SCA2, SCA3 and SCA7 patients.     

Docosahexaenoic acid induces increases in [Ca2+]i via inositol 1,4,5-triphosphate production and activates protein kinase C gamma and -delta via phosphatidylserine binding site: implication in apoptosis in U937 cells

We investigated, in monocytic leukemia U937 cells, the effects of docosahexaenoic acid (DHA; 22:6 n-3) on calcium signaling and determined the implication of phospholipase C (PLC) and protein kinase C (PKC) in this pathway. DHA induced dose-dependent increases in [Ca2+]i, which were contributed by intracellular pool, via the production of inositol-1,4,5-triphosphate (IP3) and store-operated Ca2+ (SOC) influx, via opening of Ca2+ release-activated Ca2+ (CRAC) channels. Chemical inhibition of PLC, PKCgamma, and PKCdelta, but not of PKCbeta I/II, PKCalpha, or PKCbetaI, significantly diminished DHA-induced increases in [Ca2+]i. In vitro PKC assays revealed that DHA induced a approximately 2-fold increase in PKCgamma and -delta activities, which were temporally correlated with the DHA-induced increases in [Ca2+]i. In cell-free assays, DHA, but not other structural analogs of fatty acids, activated these PKC isoforms. Competition experiments revealed that DHA-induced activation of both the PKCs was dose-dependently inhibited by phosphatidylserine (PS). Furthermore, DHA induced apoptosis via reactive oxygen species (ROS) production, followed by caspase-3 activation. Chemical inhibition of PKCgamma/delta and of SOC/CRAC channels significantly attenuated both DHA-stimulated ROS production and caspase-3 activity. Our study suggests that DHA-induced activation of PLC/IP3 pathway and activation of PKCgamma/delta, via its action on PS binding site, may be involved in apoptosis in U937 cells.