Which Tyrosine Kinase Inhibitor Must We Apply Before? A Case Report of Crizotinib-resistant Patient with Concomitant EGFR and EML4-ALK Gene Mutations

ABSTRACT The concomitant epidermal growth factor receptor (EGFR) mutation and anaplastic lymphoma kinase (ALK) translocations in lung adenocancers are very rare scenarios. Until now, 42 cases described in the literature have all been treated by different drugs. There is no overall consensus regarding the treatment for this adenocarcinoma subgroup. We report here a case of lung adenocarcinoma with concomitant EGFR mutation in exon 21 (L858R) and ALK rearrangement in primary tumour, EGFR mutation in exon 21 (L858R) and no ALK rearrangement in its synchronous metastasis. We treated this patient with crizotinib as the second-line therapy (after the first line docetaxel-cisplatin chemotherapy), but no response was obtained. The therapeutic choice for the lung adenocancer patients with concomitant EGFR mutation and ALK rearrangement is unclear. Examination of c-ros oncogene 1 mutation can be used as an indicator in the prediction of the crizotinib treatment success. The ALK mutation may not responsible for the resistance to EGFR-tyrosine kinase inhibitors (TKI), and EGFR-TKI can be initiated to EGFR and ALK dual mutant patients as the first treatment.

ALK immunohistochemistry positive, FISH negative NSCLC is infrequent, but associated with impaired survival following treatment with crizotinib.

OBJECTIVE: Metastasized non-small cell lung cancer (NSCLC) with an anaplastic lymphoma kinase (ALK) rearrangement is usually sensitive to a range of ALK-tyrosine kinase inhibitors. ALK-positive NSCLC have been identified in pivotal phase III trials with fluorescence in situ hybridization (ALK FISH+). These tumors are also expressing the fusion product (ALK immunohistochemistry (IHC)+). However, discrepant cases occur, including ALK IHC + FISH-. The aim of this study was to collect ALK IHC + cases and compare within this group response to crizotinib treatment of ALK FISH + cases with ALK FISH- cases. MATERIALS AND METHODS: In this European prospective multicenter research study patients with Stage IV ALK IHC + NSCLC treated with crizotinib were enrolled. Tumor slides were validated centrally for ALK IHC and ALK FISH. RESULTS: Registration of 3523 ALK IHC tests revealed a prevalence of 2.7% (n = 94) ALK IHC + cases. Local ALK FISH analysis resulted in 48 concordant (ALK IHC+/FISH+) and 16 discordant (ALK IHC+/FISH-) cases. Central validation revealed 37 concordant and 7 discordant cases, 5 of which had follow-up. Validation was hampered by limited amount of tissue in biopsy samples. The PFS at 1 year for ALK concordant and discordant was 58% and 20%, respectively (HR = 2.4; 95% CI: 0.78-7.3; p = 0.11). Overall survival was significantly better for concordant cases than discordant cases after central validation (HR=4.5; 95% CI= 1.2-15.9; p=0.010. CONCLUSION: ALK IHC + FISH- NSCLC is infrequent and associated with a worse outcome on personalized treatment. A suitable predictive testing strategy may be to screen first with IHC and then confirm with FISH instead of considering ALK IHC equivalent to ALK FISH according to the current guidelines.

Peripheral and bone marrow CD34+ cell levels on chronic myeloproliferative disease.

PURPOSE: The aim of the present study was to examine the relationship between peripheral CD34+ and bone marrow CD34+ levels and the clinicopathologic characteristics and laboratory parameters of myeloproliferative disease (MPD) patients. PATIENTS AND METHODS: A total of 103 MPD patients were enrolled in this study. We examined the relationship between bone marrow CD34+ and peripheral CD34+ levels and the patients' clinicopathologic and laboratory parameters. RESULTS: There were no significant correlations between the peripheral CD34+ levels and the JAK-2 V617F mutation, thrombosis, white blood cells (WBC), lactate dehydrogenase (LDH), transferrin saturation (TS), ferritin, or bone marrow cellularity. In addition, there were no significant correlations between bone marrow CD34+ levels and the JAK-2 V617F mutation, thrombosis, WBC, LDH, TS, ferritin, or bone marrow cellularity (P > 0.05). We did not identify any significant relationship between peripheral CD34+ and bone marrow CD34+ levels (P > 0.05). However, there were significant correlations between peripheral CD34+ levels and bone marrow fibrosis (P < 0.001), between bone marrow CD34+ levels and constitutional symptoms (P < 0.05), and between bone marrow CD34+ levels and bone marrow fibrosis (P < 0.001). CONCLUSION: We did not find any significant relationship between the clinicopathologic and laboratory characteristics and peripheral and bone marrow CD34+ cells from bone marrow fibrosis patients. There was also no significant relationship between bone marrow CD34+ cells and peripheral CD34+ cells. Some peripheral CD34+ cells may originate from the spleen rather than the bone marrow, which may given us different result of some parameters.

A case of multiple sclerosis and celiac disease.

Objectives. Multiple sclerosis (MS) is an inflammatory autoimmune disorder of the central nervous system (CNS). Since a correlation between gluten intake and incidence of MS had been reported, the relationship of antigliadin antibodies and MS was debated. Case Report. We report the case of a 45-year-old female MS patient who is under interferon treatment. After seven years of monitoring, during her routine gastroenterological assessment, she was diagnosed with celiac disease. Conclusion. Beside the neurological manifestations that have been demonstrated in about 10% of celiac disease (CD) patients, white-matter abnormalities in brain MRI are uncommon and controversial. But in the literature, MS seems to be associated with CD as in our patient. We suggest that MS patients with gastroenterological complaints should undergo an assessment for CD.

Diagnostic and prognostic significance of image cytometric DNA ploidy measurement in cytological samples of cervical squamous intraepithelial lesions.

OBJECTIVE: To study the DNA ploidy pattern of uterine cervical squamous intraepithelial lesions (SILs) and its diagnostic and prognostic significance. METHODS: The study included 31 cases of SIL: 11 low-grade (LSIL) and 20 high-grade (HSIL). Feulgen-pararosaniline staining was performed on previously Papanicolaou-stained smears and a DNA image cytometric study was performed. An internal reference was used to calibrate the samples. RESULTS: All 31 cases of SIL, either LSIL or HSIL, were non-diploid. Of the 11 cases of LSIL, four were tetraploid and seven were aneuploid, whereas, of the 20 cases of HSIL, four were tetraploid and 16 were aneuploid. Stemline aneuploidy was not a significant discriminator between LSIL and HSIL (P=0.32). Based on single-cell analysis, HSIL cases had significantly higher DNA content than LSIL cases (P<0.01). When a mean of 30% or more was used for the 6c-exceeding event (6cEE) value, the sensitivity and specificity to indicate HSIL were 83% and 64%, respectively, with a positive predictive value (PPV) of 81% and negative predictive value (NPV) of 65%. All HSIL cases were cervical intraepithelial neoplasia grade 2 or worse (CIN2+) on biopsy. In addition, cases which showed recurrence had more DNA content by single-cell analysis than those with an indolent clinical behaviour: P=0.04 and P=0.03 for LSIL and HSIL, respectively. CONCLUSIONS: Image cytometric DNA analysis is a useful technique for diagnostic and prognostic purposes in uterine cervical SIL when appropriate 'c' values are used in single-cell analysis. We propose that a >6c DNA content of 30% is useful as a cut-off level for predicting cases with CIN2+ in DNA image cytometry of cervical smears.

Postmastectomy radiation therapy in locally advanced breast cancer.

Despite wide efforts for early detection of breast cancer using screening mammography, locally advanced breast carcinomas account for a remarkable proportion of all breast carcinomas, particularly in developing countries. Locally advanced breast cancer may have widely different clinical and biological features. Radiotherapy plays an important role in the management of locally advanced breast cancer. Postmastectomy radiotherapy has been shown to significantly reduce the risk of loco-regional failure and to improve disease free survival as well as overall survival in high-risk women with breast cancer. This review article summarizes the data from randomized trials revealing a significant benefit from postmastectomy radiation therapy in patients with locally advanced disease.

Hypoxia inducible factor-1 alpha and carbonic anhydrase IX overexpression are associated with poor survival in breast cancer patients.

PURPOSE: Hypoxia is common in many solid tumors such as breast, head-neck, and soft tissue malignancies. Hypoxia causes overexpression of hypoxia inducible factor-1 alpha (HIF-1α) and carbonic anhydrase IX (CA IX) which are associated with unfavorable prognosis in breast cancer. In our study, we evaluated HIF-1α and CA IX expression in patients with breast cancer. METHODS: Between June 1996 and June 2008, 111 women with breast cancer were evaluated. Estrogen receptor (ER) and progesterone receptor (PR) status and Her2/ neu expression were evaluated by immunohistochemical methods. Her-2/neu expression was also assessed by FISH method when needed. Two groups were created: ER and PR positive, Her-2/neu negative (group 1, n=56); and ER and PR negative, Her-2/neu positive (group 2, n=55). HIF-1α and CA IX expressions were investigated in both groups and results were compared. In addition, we investigated the association between HIF-1α and CA IX expressions with stage, grade, lymph node metastasis, tumor size, menopause status and survival. RESULTS: Median patient age in group 1 was 52 years (range 34-77), and in group 2 47 years (range 27-83). HIF-1α expression was detected in 26 (46.4%) of group 1 and in 46 (83.6%) of group 2 patients (p=0.0001). CA IX expression was detected in 25 (46.4%) of group 1 and in 37 (67.3%) of group 2 patients (p7equals;0.0137rpar;. In group 1, median disease free survival (DFS) was 97 months and in group 2 46 months (p=0.0308). In group 1, median overall survival (OS) was 108 months and in group 2 75 months (p=0.0339). CONCLUSION: HIF-1α and CA IX overexpressions are observed more often in ER and PR negative, Her-2/neu positive breast cancer and are associated with poor survival.

The impact of bone marrow fibrosis on the outcome of hematopoietic stem cell transplantation.

We retrospectively analyzed the data of 175 patients who underwent autologous (n = 69) or allogeneic hematopoietic stem cell transplantation (HCT) (n = 106) including 19 (27.5%) and 38 (35.8%) recipients who had bone marrow fibrosis (BMF) prior to transplantation, respectively. We investigated the effects of BMF on engraftment, graft-versus-host disease (GVHD), early posttransplant complications, and survival. Pretransplantation BMF did not delay engraftment and showed no impact either on early posttransplant complications or on the development of acute and/or chronic GVHD. Probability of 1-year overall survival (OS) and progression-free survival (PFS) of autologous HCT recipients were similar, namely 76.7% versus 88.6% (P > .005) and 26.33% versus 16.5% (P > .05) among patients with versus without fibrosis, respectively. In allogeneic HCT recipients, the probability of 1-year OS was 35.2% among patients with versus 48.9% among those without fibrosis (P = .004) PFS at 1 year was inferior among allogeneic HCT recipients with BMF: 27.8% versus 51.2% (P = .0008). Cox regression analysis revealed BMF to be independently associated with age, Sorror comorbidity index, primary disease, and disease status during HCT (P = .045).

Ectopic primary B cell lymphoma of the thyroid presenting as an anterior mediastinal mass. A case report.

Ectopic thyroid tumours arising in the mediastinum without connection to the cervical thyroid gland are very rare. Follicular adenoma, papillary carcinoma and follicular carcinoma in the mediastinum has been reported, but primary ectopic thyroid B cell lymphoma has not been reported previously. We report mediastinal primary ectopic thyroid large B cell lymphoma in an 80-year-old man. Differential diagnosis from primary mediastinal large B cell lymphoma and clinicopathologic features are discussed.

Vascular endothelial growth factor, c-erbB-2 and c-erbB-3 expression in colorectal adenoma and adenocarcinoma.

AIM: To analyze vascular endothelial growth factor (VEGF), c-erbB-2 and c-erbB-3 expression and to evaluate their relation to clinicopathologic parameters and pathogenesis of colorectal carcinoma. METHODS: Sections of adenoma, intramucosal carcinoma and adenocarcinoma were evaluated by immunohistochemistry in 85 malignant and 37 benign colorectal neoplasms for the expression of VEGF, c-erbB-2 and c-erbB-3 considering clinicopathological variables. RESULTS: VEGF was detected in comparable percentages of all neoplasm types while c-erbB-2 expression was detectable more frequently in adenoma than adenocarcinoma cases (65% vs 43%). Except for the correlation of c-erbB-3 expression with Dukes' staging, there was no correlation between the studied markers and grade of differentiation, Dukes' stage and localization of colorectal adenocarcinoma. c-erbB-3 expression was seen more frequently in tubular adenomas, while c-erbB-2 expression was higher in tubulovillous and villous types. These differences were not statistically significant. The presence of distant metastasis and angiolymphatic invasion were identified as independent predictors of survival. A positive correlation was found between VEGF expression and lymphatic vessel invasion and regional lymph node involvement. CONCLUSION: These results suggest that VEGF, c-erbB-2, c-erbB-3 expression does not have prognostic value in colorectal cancer. VEGF expression may be implicated in the lymph node metastasis.

Candida vertebra osteomyelitis in a girl with factor X deficiency.

Candidal vertebra osteomyelitis is a rare condition which occurs primarily in immunocompromised patients. We report a 14-year-old girl with factor X deficiency who developed candida vertebra osteomyelitis during home therapy. The microorganism was probably from a contaminated peripheral cannula used for infusion of factor concentrate. This is the first such case in bleeding disorders to our knowledge.

A short duration transient ischemia induces apoptosis in retinal layers: an experimental study in rabbits.

PURPOSE: To investigate retinal cell apoptosis in an experimental transient, short duration ocular ischemia model. METHODS: An experimental ischemia model, which simulates creating temporary high intraocular pressure to control intraocular bleeding during pars plana vitrectomy, was set up. Rabbits were randomly divided into three groups. Group 1 was the control group. In Group 2, intraocular pressure was increased to 97 mmHg for 5 minutes. In Group 3, intraocular pressure was increased to 97 mmHg for 10 minutes. After 24 hours, terminal deoxyribonucleotidyl transferase-mediated dUTP-digoxigenin nick-end labeling assay was used to detect retinal apoptosis in rabbit eyes. Only nuclear staining in retinal cells was counted. RESULTS: Groups with 5 minutes and 10 minutes of ischemia showed significantly higher amount of ganglion cell layer apoptosis when compared with the control group (p<0.05). Light microscopy and standard hematoxylin-eosin did not show any significant damage in the retina cells. CONCLUSIONS: Apoptotic cell death in the retinal cell layers occurs in temporary ischemia-reperfusion as early as 5 and 10 minutes duration.

Primary omental pregnancy: case report.

Omental pregnancy is a very rare form of ectopic pregnancy. Here we presented a case of primary omental pregnancy diagnosed at surgical exploration. A 28 year old woman submitted with severe abdominal pain, without any delay of menstruation. History of the patient revealed no use of contraceptive method. There was no gestational sac in the endometrial cavity and no tubal ring in the adnexa, but free peritoneal fluid in the pouch of Douglas was detected at ultrasonography. Laparotomy was done according to pre-operative diagnosis of ruptured tubal pregnancy. Bilateral tubes and ovaries were intact; omental pregnancy was detected and partial omentectomy was performed. Although 16 cases of omental pregnancy (mostly secondary) were reported in the literature, herein we describe a primary omental pregnancy without adnexial involvement.

P21 and Bax expression in cutaneous malignant melanomas: correlation with histologic prognostic parameters.

In response to DNA damage, p53 accumulates and regulates expression of several genes, including cyclin-dependent kinase inhibitor p21. Cells then undergo p21 dependent cell cycle arrest, which allows DNA damage repair and apoptosis. Bax is a death promoter member of the bcl-2 family which plays a central role in the regulation and commitment to programmed cell death. Breslow thickness is the most important factor in predicting prognosis for cutaneous malignant melanoma. In order to define the role of cyclin dependent kinase inhibitors and apoptosis regulators in invasion of malignant melanoma we investigated the expression of p21 and bax proteins. We observed that significant high p21 expression was associated with increasing Breslow thickness (Spearman correlation analysis, p=0.01). Additionally, Clark level I and II tumours expressed significantly lower p21 positivity than Clark level III, IV and V (p=0.006). Similarly, thick tumors showed a higher bax expression (p=0.012). Our results suggested that the role of p21 expression is more complicated in melanocytic skin cancers and abnormal regulation or abnormal function of cell cycle regulators occurred in the development and progression of malignant melanoma. In order to understand the role of bax expression in thick malignant melanomas and invasion biology, comparative analytic studies with other apoptosis regulators are needed.

Inhibition of epidural scar tissue formation after spinal surgery: external irradiation vs. spinal membrane application.

PURPOSE: The scar tissue that forms after lumbar dissection is a severe complication and a cause of lumbar and radicular pain. It was recently shown that radiotherapy could inhibit peridural fibrosis after laminectomy. In this study, the efficiency of external irradiation was compared with spinal membrane application. METHOD AND MATERIALS: Thirty male New Zealand rabbits underwent L5 laminectomy. Ten rabbits each received a single fraction of 900-cGy external irradiation administered by 9-MeV electron beam 24 h after the surgery. Ten other rabbits each had spinal membrane applied during laminectomy. The remaining 10 rabbits constituted the control group. All of the rabbits were killed 30 days after the laminectomy. Axial histologic sections through the laminectomy defect were evaluated. Each specimen was scored for the extent and density of fibrosis and arachnoidal adherence. RESULTS: The extent and density of fibrosis and arachnoidal adherence differed significantly between the control group and the treatment groups (p < 0.05). However, the extent and density of fibrosis and arachnoidal adherence did not differ significantly between the spinal membrane and irradiation groups (p > 0.05). CONCLUSION: This preliminary study showed that high-single-fraction/low-total-dose administered postoperatively can successfully inhibit postsurgical epidural fibrosis as effectively as applied spinal membrane.

Association between the severity of heart failure and the susceptibility of myocytes to apoptosis in patients with idiopathic dilated cardiomyopathy.

BACKGROUND: Bcl-2 proto-oncogene, an inhibitor of apoptosis and Bax proto-oncogene, an inducer of apoptosis play critical roles in the molecular circuit controlling apoptosis in cardiac muscle. The ratio of Bax to Bcl-2 proto-oncogene determines survival or death after an apoptotic stimulus. We speculated that susceptibility of myocytes to apoptosis determined as the Bax/Bcl-2 ratio might vary with the severity of heart failure. METHODS AND RESULTS: We studied immunohistochemically 108 endomyocardial biopsy specimens from 30 patients with idiopathic dilated cardiomyopathy (mild heart failure, n=14; moderate or severe heart failure, n=16) with the use of Bcl-2 and Bax monoclonal antibodies. The expression of each protein was determined semiquantitatively as the fraction of myocytes labeled with specific monoclonal antibodies using a digital morphometric analysis system. Patients with mild heart failure showed significantly increased Bax/Bcl-2 ratio than the patients with advanced heart failure (1.59+/-1.26 vs. 0.34+/-0.43, P=0.002). The expression of Bcl-2 was found to be independent of the severity of heart failure whereas the expression of Bax was significantly higher in patients with mild heart failure compared to the patients with moderate or severe heart failure (52.1+/-29.3 vs. 21.6+/-22.4%, P=0.005). Additionally, Bax/Bac-2 ratio was inversely correlated with the mitral E-interventricular septum distance, left ventricular end-systolic and end-diastolic diameter. CONCLUSION: The susceptibility of myocytes to apoptosis is significantly increased in the early phase of heart failure but it decreases with worsening of the disease due to depressed expression of Bax onco-protein. Increased myocyte susceptibility to apoptosis may have a role in the transition from mild heart failure to severe in patients with idiopathic dilated cardiomyopathy.