Caspase-8 activation by TRAIL monotherapy predicts responses to IAPi and TRAIL combination treatment in breast cancer cell lines.

The discovery of cancer cell-selective tumour necrosis factor-related apoptosis inducing ligand (TRAIL)-induced apoptosis generated broad excitement and development of TRAIL receptor agonists (TRA) as potential cancer therapy. Studies demonstrating the synergistic combination effect of SMAC mimetics and TRA further suggested potentially effective treatment in multiple tumour settings. However, predictive biomarkers allowing identification of patients that could respond to treatment are lacking. Here, we described a high throughput combination screen conducted across a panel of 31 breast cancer cell lines in which we observed highly synergistic activity between TRAIL and the inhibitors of apoptosis proteins (IAP) inhibitor (IAPi) AZD5582 in ~30% of cell lines. We detected no difference in the expression levels of the IAPi or TRAIL-targeted proteins or common modulators of the apoptotic pathway between the sensitive and resistant cell lines. Synergistic combination effect of AZD5582 and TRAIL correlated with sensitivity to TRAIL, but not to AZD5582 as a single agent. TRAIL treatment led to significantly greater activity of Caspase-8 in sensitive than in resistant cell lines (P=0.002). The majority (12/14) of AZD5582+TRAIL-resistant cell lines retained a functional cell death pathway, as they were sensitive to AZD5582+TNFα combination treatment. This suggested that failure of the TRAIL receptor complex to transduce the death signal to Caspase-8 underlies AZD5582+TRAIL resistance. We developed a 3D spheroid assay and demonstrated its suitability for the ex vivo analysis of the Caspase-8 activity as a predictive biomarker. Altogether, our study demonstrated a link between the functionality of the TRAIL receptor pathway and the synergistic activity of the IAPi+TRA combination treatment. It also provided a rationale for development of the Caspase-8 activity assay as a functional predictive biomarker that could allow better prediction of the response to IAPi+TRA-based therapies than the analysis of expression levels of protein biomarkers.

Transarticular approach and retrograde plate osteosynthesis (TARPO) using implants with angular stability -- a series of 17 cases of complex distal femoral fractures type C3/AO.

AIM: The aim of this retrospective study was to evaluate the outcome of distal femoral fractures type C3 AO, using TARPO technique and plates with angular stability. MATERIAL AND METHODS: The study included 17 fractures type C3 AO, with 4 open fractures: 1 type I, 1 type II, and 2 type IIIA with bone loss Gustilo. All patients were operated by TARPO technique using Less Invasive Stabilization System-LISS (4 cases), Locked Compression Plates-LCP (8 cases) and plates with polyaxial stability (5 cases). The excellent stability of the construct allowed fast knee rehabilitation. The follow-up period included at least 12 months. RESULTS: 15 fractures healed within a mean time of 12.6 weeks,while 2 cases with open fractures and bone loss required secondary bone grafting. We recorded no infection or implant failures. The outcome using Neer scale was excellent in 9 cases and satisfactory in 7 cases (1 patient with discontinued follow-up). CONCLUSIONS: This demanding TARPO technique has the advantage of a faster rate of union and improved exposure of the knee joint. The locked plates provide a unique alternative in distal femoral fractures type C3 AO, as well as in osteoporotic and open fractures.

Surgical approach in difficult tibial pilon fractures.

AIM: The aim of the present study is to emphasize the optimal moment and approach for tibial pilon complex fractures and their complications. MATERIAL AND METHODS: We have investigated and treated 7 patients with complex fractures of the tibial pilon with tendencies to complications. According to AO classification,we have recorded 4 fractures type 43C2 AO, 2 fractures type 43C3 AO (with one open fracture type IIIA Gustilo) and 1 fracture type 43B3 AO. According to soft tissue status, surgical treatment was performed by a one-step or two-step approach. RESULTS: All fractures healed after an interval of 13.6 weeks(range 8-28 weeks) with one delayed consolidation. The open fracture was complicated by posttraumatic arthrosis. No infections or implant failures were recorded. According toOvadia score, objective and subjective evolution showed good results in 5 cases, one excellent result and one poor result. CONCLUSIONS: Fracture type and the soft tissue status are crucial for the results of operated fractures. These arguments determine the application of one-step or two-step surgical procedure as well as optimal implant choice.

E-cadherin expression in invasive ductal carcinoma associates ultrastructural changes in desmosomes structure.

UNLABELLED: The relationship between E-cadherin presence/absence and the integrity of the desmosomes at the same level becomes important regarding the discrepancies between E-cadherin lack of expression in severe invasive carcinomas and the desmosome activity. PURPOSE: In the present study, we have evaluated the presence of E-cadherin (EC) in 34 cases of metastatic or non-metastatic invasive ductal breast carcinomas (IDC) by immunohistochemistry followed by transmission electron microscopy (TEM) evaluation on samples prepared from paraffin sections. MATERIALS AND METHODS: Our study has analyzed 34 paraffin blocks incoming from 20 cases with documented presence of metastatic invasion and 14 cases without metastases. All samples were processed and stained by classic Hematoxylin-Eosin, immunohistochemistry to detect EC presence and transmission electron microscopy. RESULTS: Our results, even on a small pilot group, emphasize that EC presence is associated with the complete desmosomal integrity in non-metastatic cases, at least for the investigated areas. From the metastatic IDC cases, we have observed reduced EC expression in only three cases and important loss of desmosomal arrangement, mainly at the desmosomal plate level. CONCLUSIONS: This observation can issue the hypothesis that even in IDC cells that express EC, the invasive potential depend not only on EC-dependent junctions but also on the desmosome integrity. Also, correlated relationship between EC expression, potentially explored desmogleins, desmocollin, desmoplakin and plakoglobin expression and TEM ultrastructure can lead to a conclusion about the invasive potential of these malignant cells.

Trigeminal nerve: MRI anatomy and case presentation of trigeminal neuralgia due to arterial compression.

Trigeminal neuralgia (TN), also known as tic douloureux is a chronic neuropathic pain disorder characterized by sporadic episodes of extreme, sudden burning or shock-like face pain that last from a few seconds to 2 minutes. Trigeminal neuralgia has a reported incidence of 5.9/100,000 women and 3.4/100,000 men in USA. The exact pathophysiology is still unclear, but demyelization leading to abnormal discharge in fibers of the trigeminal nerve is a probable cause. In the majority of cases, no structural lesion is detected but in almost 15% of patients medical imaging methods like MRI, CT or angiography can identify a vein or artery that compresses the nerve which results in focal demyelization. The authors present a case of trigeminal neuralgia investigated by MRI, which identified a vascular compression of the nerve 9 mm after emerging the pons by the superior cerebellar artery (SCA) and one of its branches. The authors also realize a review of the MRI anatomy of the trigeminal nerve.

Electron microscopy analysis of skin biopsies in CADASIL disease.

Cerebral Autosomal Dominant Arteriopathy with Subcortical Infarcts and Leukoencephalopathy (CADASIL) is an inherited vascular disorder, non-amyloid and non-atherosclerotic, affecting predominantly the central nervous system. We examined samples of skin biopsies from six patients (men, 43-52-year-old), admitted for treatment in the Neurology Clinic regarding the presence of partial motor impairment on upper and lower right limbs, facial asymmetry and phrasing impairment (three of the patients); These three patients had family history remarkable for early-onset strokes: mother and two brothers deceased by early strokes (40-50-year-old). Skin biopsy samples were fixed in glutaraldehyde and post-fixed in osmium tetroxyde. After dehydration, tissue samples were embedded in Epon. Ultrathin sections were mounted on copper grids and stained with uranyl acetate and lead citrate as usual and examined with a transmission electron microscope Phillips CM100. In all cases ultrastructural study showed granular osmiophilic material (GOM) in extracellular locations, between degenerating smooth muscle cells in dermal arteries or in their indentations. Deposits of GOM varied in size and electron density. Degeneration and loss of smooth muscle cells (SMCs) leads to abnormal enlargement of the space between these cells Ultrastructural analysis in three cases showed chromatin condensation and peripheral aggregation of nuclear material suggesting cells entry to apoptosis. These aspects and the marked destruction of the vascular wall were correlated with MRI findings and the severity of clinical manifestations at these patients. Our study showed that findings of GOM deposits, degeneration and loss of SMCs (probably by apoptosis), cell adhesion elements disturbance are characteristic for CADASIL disease and sufficient for diagnose of certainty. Moreover, electron microscopy analysis of skin biopsies is a useful tool for a differential diagnosis and can be considered as first choice method.

[Ultrastructural aspects in the development of the human nephron--electron microscopy study]. / Aspecte ultrastructurale în dezvoltarea nefronului uman. Studiu de microscopie electronica.

UNLABELLED: We have looked for electron microscopy aspects tissue fragments of human nephrons harvested from fetuses of 7,5-19 weeks to show ultrastructure aspects of the nephron during development. MATERIAL AND METHODS: We have used 9 fragments from human embryos; Four cases were aged of 7,5, 8, 12 and 17 weeks. The other 5 cases were between 18 and 19 weeks. Tissue fragments were fixed in glutaraldehyde at 4 degrees C and processed by the classic technique. RESULTS: The electron microscopy study shows the evolutive steps of the human nephron (stages I-IV), stages met for the 12-19 weeks embryo. For the cases under this age the meta-nephrogenic blastema was dominant. In the IVth development stage, the cells in the proximal tubule show a higher development degree than the distal segments. CONCLUSION: These observations indicate that during early development the proximal segment is better developed and probably more functional than the distal segments.

[The influence of the dihydroxamic compounds on the rat lung]. / Influenta unor compusi dihidroxamici asupra plamanului de sobolan.

The role of the dihydroxamic compounds in the human and animal organism are little known. This is the reason they the experiment studies the effect of these compounds on the rat lung. 3 lots of Wistar adult male rats were used. The first lot was the witness lot and did not receive any substance. The second lot received Dihydroxamic adipoil acid, 0.5 mg/kg weight/ day, intraperitoneal administration in a unique daily dose, for 3 weeks. The third lot received Dihydroxamic terephtaloyl acid, 0.5 mg/kg weight/day, intraperitoneal administration in a unique daily dose. After 14 and 21 days the animals of each lot were euthanised by sectioning carotid arteries. The authors collected the lungs, prepared the histological sections, stained in HE and Szekely. The witness lot presented normal rat lung structure. After 2 and 3 weeks of treatment, the second lot presented distended blood vessels. After 2 and 3 weeks of treatment, the third lot presented very severe alterations of the lung tissue at all levels.

[Surgery experience regarding the pathology of the greater omentum]. / Morfopatologia omentumului mare chirurgical.

This study followed the evaluation of the surgical pathology of the greater omentum. From the casuistry of Urology Surgery Clinic in Hospital 2 Iasi, we have selected 1500 omentum pathologic specimens (inflammatory, traumatic, tumoral) that were investigated macroscopically and microscopically. The specimens were harvested during 1993-2000. Acute inflammatory involvement of the greater omentum recognizes a primitive etiology in 8-10% cases and secondary one in 90-92% cases; these lesions are associated to some inflammatory processes of peritoneal viscera by direct or indirect mechanisms. Non-tumoral pathology was signaled in 201 cases, malignant and benign tumors were found in 198 cases. Acute catarrhallis and phlegmonous omentitis were identified in: peritonitis by digestive perforation (240 cases), bladder perforation (12 cases), broken pyosalpinx (18 cases), acute calculosic colecystitis (260 cases), hernia (75 cases), strangled eventration (65 cases). The conclusion of our study is that there is a specific pathology for the omentum but also the latter is largely involved in the emergency and iterative abdominal surgical pathology.

[Gelatinases in peri-implantation stage]. / Gelatinazele în perioada periimplantara.

The periimplantation stage in mice is marked around the 5th day after mating by blastocyst presence in the emdometrium. The events that are marking the implantation event are focused on the evolution of the extracellular matrix around the mezenchime cells that undergoes decidualization. The inductive process is guided by gelatinases and the purpose of our study is to demonstrate their presence at this level.

Actinic keratosis--a histoenzymological study.

Actinic keratosis or senile keratosis is the most frequent premalign epithelial lesion that occurs in the elderly with a history of prolonged and intense exposure to the UV radiation and with an inborn susceptibility. Although the genetics and risk factors are clear nowadays, the histogenesis of this lesion is still under study. The histoenzymologic tests (such as those that demonstrate the ATP-ase and SDH-ase activity) are very useful for early detection of the structural and biochemical changes within the actinic keratosis lesion.

[Periodontitis, a risk factor for cardiovascular diseases?]. / Periodontitele, factor de risc pentru bolile cardiovasculare?

Exist a statistically significant association between a diagnosis of coronary heart disease and certain oral health parameters. These parameters are independent and more strongly associated with coronary heart disease than were recognized risk factors, such as serum cholesterol levels, body mass index, diabetes and smoking status. The correlation between serum and salivary cholesterol concentration and the role of salivary lipids in oral health have been poorly characterized. Lipids in saliva are mostly glandular in origin, through some may diffuse diffuse directly from the serum. The surface of the enamel (in case of dietary fat which have an cariostatic effect) is protected against demineralization by a mixture of salivary lipids and proteins. Salivary lipids may have anucleating role in the early mineralization of the dental plaque. The prevalence of periodontal disease may be increased in patients with atherosclerosis, and dental and periodontal infections have been shown to be associated with occurrence of acute and chronic coronary heart disease. Among the known risk factors of atherosclerosis, high serum lipid levels, especially cholesterol may be important with regard to periodontal health.

Ultrastructural aspects of the alveolar wall in SIDS.

The aim of this study is to make a comparative analyse between light and ultrastructural lesions of the alveolar wall in SIDS. The study was made on lung specimens collected from autopsy of infants with sudden unexplained death. There were studied histopathologically 10 cases of sudden death. Only in 3 cases were made electron microscopically studies. At light microscope, there were put in evidence interstitial and intra-alveolar oedema and the presence of many parietal and intra-alveolar macrophages. There is a good correlation between light and ultrastructural aspects.

New trends in "mystery SIDS" research.

The aim of this study is to suggest some new trends of research in SIDS, leaving from the pathologist's observations of the presence of unusual lesions in SIDS. The histopathological study was made on 27 cases of SD, representing 1996 SIDS cases registered to hospital. There were put in evidence the oesophageal inflammation, the hyperplasia of submucosal laryngeal glands, different grades of bronchiolar mucous hypersecretion and reducing of elastic lung tissue and the hypertrophy of muscular lung small arteries. This lesions are frequently encountered to SIDS cases.