Mariamma Joseph

Comparison of Human Telomerase Reverse Transcriptase Messenger RNA and Telomerase Activity as Urine Markers for Diagnosis of Bladder Carcinoma

AbstractBackground: Human telomerase reverse transcriptase (hTERT) has been identified as the catalytic subunit of telomerase ribonucleoprotein complex known to be required for cellular immortality and oncogenesis. Although human telomerase activity (hTA) is considered as a general marker for malignancy based on its presence in most malignant tumors including bladder cancer, its detection in urine is affected by many factors. The objective of this study was to compare the clinical utility of detecting urine hTERT messenger RNA (mRNA) by multiplex hTERT/GAPDH RT-PCR and urine hTA by telomerase repeat amplification protocol (TRAP) in the diagnosis of bladder cancer. Methods and Results: Cystoscopy urine samples or bladder washes prospectively collected from 35 patients with confirmed (35) or clinically suspected (5) transitional cell carcinoma (TCC) of the bladder were examined by TRAP, hTERT/ GAPDH RT-PCR, and urine cytology. The control group comprised 21 healthy volunteers and 3 patients without TCC. The hTERT/GAPDH RT-PCR test showed significantly higher diagnostic sensitivity than TRAP assay (94.3% vs 48.6%, P <.001) and urine cytology (95.2% vs 61.9%, P =.008) for confirmed TCCs. In particular, for superficial TCCs low grade (I–II), the hTERT/GAPDH RT-PCR test outperformed TRAP (90% vs 25%, P <.001) and urine cytology (91.7% vs 58.3%, p =.46). The overall specificity of the hTERT/GAPDH RT-PCR, TRAP and urine cytology was 92% (22/24), 100% (24/24), and 100% (3/3), respectively. A positive hTERT mRNA expression was also detected in urologic specimens from 3 patients with previous history of TCC, 3 to 6 months before cystoscopic evidence of cancer. Conclusion: In this pilot study, the hTERT mRNA expression in urine sediments is a more sensitive marker for diagnosis of TCC of the bladder than hTA and cytology. However, there is a higher false-positive rate.

Role of placental metallothionein in maternal to fetal transfer of cadmium in genetically altered mice

The role of placental metallothionein (MT) as a barrier for maternal to fetal transfer of cadmium (Cd) was investigated using mice which overexpressed the MT-1 isoform (MT-1*), mice which did not express the MT-1 and 2 isoforms (MT-null) and control mice (C57BL/6). In addition, immunohistochemical localization of MT in the placenta was determined in these mice. Two days prior to parturition, the mice were injected with radioactive 109Cd chloride (4 microCi, 0.6 ng Cd/mouse) and killed 24 h later. Organs and fetuses were collected and radioactivity, MT and metal levels were measured. Cd accumulated mainly in the liver and kidney (80% of administered dose) with very low levels (0.1-0.3%) detected in fetuses. When analyzed on a per organ or per gram basis, MT-null fetuses accumulated significantly more Cd (3-10-fold) than the control fetuses and there was no significant difference in fetal Cd accumulation in the MT-1* and control fetuses. As expected, MT and zinc levels were higher in MT-1* than C57BL/6 mice and no MT was detected in MT-null mice. Most striking was the high hepatic MT levels in MT-1* dams (4 mg/g). Immunohistochemical analysis showed MT staining in spongiotrophoblasts, glycogen cells, visceral yolk sac, trophoblast giant cells and maternal decidual cells with the MT-1* placenta staining much more intensely as compared to control placenta. The results suggest that placental MT reduces maternal to fetal Cd transfer, however the low doses of Cd administered in the present experiment resulted in high levels of Cd accumulation in liver and kidney in all groups of mice with a low concentration of Cd reaching the placenta. Thus, the role of placental MT as a barrier for Cd is inconclusive.

Topical Imiquimod Therapy for Lentigo Maligna

Lentigo maligna (LM) presents a challenge for complete surgical excision. Imiquimod is a topical immune-response modifier that acts on the immune system. We report our experience using imiquimod 5% cream as a surgical alternative for treatment of LM. Consecutive patients between December 2004 and February 2006 with LM were treated with topical imiquimod. Data on patient and lesion characteristics, side effects of therapy, posttreatment biopsy results, and follow-up was collected. Seven patients were treated with imiquimod 5 nights/wk for 12.4 weeks. Complete histologic and clinical resolution was seen in 86% (6 of 7 patients), at 19.1 months follow-up. Side effects included erythema (86%) and crusting (71%), resulting in dose alteration in 71% of patients. Topical imiquimod therapy demonstrates a high response rate for treatment of LM, with tolerable side effects. Further investigation into its efficacy in the treatment of LM in controlled clinical trials is warranted.

Atypical squamous cells of undetermined significance: A cytohistological study in a colposcopy clinic.

Cytohistologic correlation was performed by 3 observers on 100 atypical squamous cells of undetermined significance (ASCUS) cases from a colposcopy clinic. Our objectives were to: 1) subclassify ASCUS cases and determine their clinical significance; 2) assess the independent predictive value of different cytologic parameters for biopsy‐proven dysplasia (BPD); and 3) calculate interobserver variability. The prevalence of BPD was 73% in the ASCUS favor dysplasia (AFD) group, and 27% in the ASCUS favor reactive (AFR) group (P  0.001). The sensitivity of cervical cytology (AFD) for detecting BPD was 88.5%. Using multiple logistic regression, only nuclear membrane irregularity was found to be independently predictive of BPD (P  0.0001). The interobserver agreement (kappa) was 0.41. Colposcopic smears were inferior to referring smears in detecting dysplasia, with 67% of patients having a referring diagnosis of dysplasia. In a colposcopy clinic population there is a high prevalence (73%) of BPD in the AFD group. Attention should be paid to nuclear membrane irregularity in the risk stratification of ASCUS cases. Diagn. Cytopathol. 1999;21:211–216.

Obtaining high cure rates for challenging facial malignancies: A new method for producing rapid, accurate, high-quality frozen sections

PURPOSE The authors developed a new system to provide rapid, accurate, full-face frozen sections. OBJECTIVE To evaluate the efficacy of the system when applied to the treatment of nonmelanoma cutaneous malignancies using Mohs micrographic surgery (MMS). METHODS Patients undergoing MMS procedures between 2003 and 2007 for nonmelanoma head and neck cutaneous malignancies were prospectively collected. Specimens were prepared either in a traditional cryostat-based manner or using the new system. RESULTS A total of 196 patients with 234 head and neck nonmelanoma cutaneous malignancies were included. The majority of tumours were basal cell carcinomas (89.5%). Of these, 38% demonstrated aggressive histologies (sclerosing or micronodular), and 30% were recurrent. On average, two levels (range one to six) and four blocks (range two to 23) were required to obtain clear margins. The mean defect size was 3.68 cm(2) (range 0.13 cm(2) to 37.68 cm(2)). Over the five-year study period, there were two recurrences in 234 cases (less than 1%), which compares favourably with other MMS series. The new system was associated with a shorter operative time than traditional specimen preparation (102 min versus 131 min; P=0.004). The new and traditional specimen preparation groups were similar in terms of the number of previous recurrences (29% versus 30%; P=1.00), defect size (3.7 cm(2) versus 4.0 cm(2); P=0.81) and the number of levels required (1.9 versus 1.5; P=0.05). CONCLUSIONS The new system enables fast, accurate, full-face frozen section specimens that are ideal for MMS. The speed of specimen preparation is demonstrated by faster operative times, and a low recurrence rate attests the accuracy and quality of the sections.

A Freezing Technique for Base Mold Embedding of Multiple Block Frozen Sections

Abstract This article describes a technique for preparing multiple frozen section blocks outside the cryostat chamber. Specimens are first oriented in modified disposable base molds (“platform molds”) in a process similar to paraffin embedding. Freezing takes place in a “CryoCaddy,” which consists of a freezing platform cooled with dry ice and housed in an insulated container. During freezing, the mold is flooded with a tissue fieezing compound, and a chuck is joined to the mold. This establishes a parallel relationship between the chuck, block face, and cutting plane of the cryostat, which ensures that minimal trimming-in is required quired before full-faces ections are obtained. This technique successfully avoids specimen compression and distortion (crush artifact) that may be produced when a weighted heat extractor is used. Furthermore, this technique eliminates ice crystal artifact that occurs when freezing proceeds too slowly. The CryoCaddy has sufficient thermal inertia to allow the preparation of multiple blolcks over several hours, thereby allowing the cryostat to be dedicated to cutting. This fast, simple, and reproducibly accurate technique is ideal for frozen section evaluation of multiple block specimens and/or when precise orientation is required. (The J Histotechnol 30:23, 2007) Submitted May 23, 2006; accepted with revisions November 9, 2006

Comprehensive gene expression profiling identifies distinct and overlapping transcriptional profiles in non-specific interstitial pneumonia and idiopathic pulmonary fibrosis

BackgroundThe clinical-radiographic distinction between idiopathic pulmonary fibrosis (IPF) and non-specific interstitial pneumonia (NSIP) is challenging. We sought to investigate the gene expression profiles of IPF and NSIP vs. normal controls.MethodsGene expression from explanted lungs of patients with IPF (n = 22), NSIP (n = 10) and from normal controls (n = 11) was assessed. Microarray analysis included Significance Analysis of Microarray (SAM), Ingenuity Pathway, Gene-Set Enrichment and unsupervised hierarchical clustering analyses. Immunohistochemistry and serology of proteins of interest were conducted.ResultsNSIP cases were significantly enriched for genes related to mechanisms of immune reaction, such as T-cell response and recruitment of leukocytes into the lung compartment. In IPF, in contrast, these involved senescence, epithelial-to-mesenchymal transition, myofibroblast differentiation and collagen deposition. Unlike the IPF group, NSIP cases exhibited a strikingly homogenous gene signature. Clustering analysis identified a subgroup of IPF patients with intermediate and ambiguous expression of SAM-selected genes, with the interesting upregulation of both NSIP-specific and senescence-related genes. Immunohistochemistry for p16, a senescence marker, on fibroblasts differentiated most IPF cases from NSIP. Serial serum levels of periostin, a senescence effector, predicted clinical progression in a cohort of patients with IPF.ConclusionsComprehensive gene expression profiling in explanted lungs identifies distinct transcriptional profiles and differentially expressed genes in IPF and NSIP, supporting the notion of NSIP as a standalone condition. Potential gene and protein markers to discriminate IPF from NSIP were identified, with a prominent role of senescence in IPF. The finding of a subgroup of IPF patients with transcriptional features of both NSIP and senescence raises the hypothesis that “senescent” NSIP may represent a risk factor to develop superimposed IPF.

Successful Treatment of Fibrosing Organising Pneumonia Causing Respiratory Failure with Mycophenolic Acid.

Organising pneumonia (OP) is usually promptly responsive to corticosteroid treatment. We describe a series of 3 cases of severe, progressive, biopsy-proven fibrosing OP causing respiratory failure. All cases presented with peribronchial and subpleural consolidations, had a fibro-inflammatory infiltrative component in the alveolar septa, and only had a partial and unsatisfactory response to corticosteroids. However, they responded to mycophenolic acid (MPA) treatment with resolution of respiratory failure as well as clinical and functional improvement. MPA as an additional treatment option for aggressive forms of fibrosing OP and interstitial lung disease needs to be further explored.

Endobronchial Carcinoid Tumour with Extensive Ossification: An Unusual Case Presentation

Carcinoid tumour is a well-known primary endobronchial lung neoplasm. Although calcifications may be seen in up to 30% of pulmonary carcinoid tumours, near complete ossification of these tumours is an unusual finding. Such lesions can prove diagnostically challenging at the time of intraoperative frozen section as the latter technique requires thin sectioning of the lesion for microscopic assessment. We present an unusual case of endobronchial carcinoid tumour with extensive ossification in a 45-year-old male. Preliminary intraoperative diagnosis was achieved through the alternative use of cytology scrape smears. The final diagnosis was confirmed after decalcification of the tumour. The prognostic implications of heavily ossified carcinoid tumours remain elusive. Long-term clinical follow-up of these patients is recommended.

Vasculitis masquerading as drug allergy: thinking outside the ‘adult’ box of possible diagnoses

Case report A 32 year-old male presented with fever and pharyngitis. Amoxicillin was prescribed and 5 days into therapy he developed a petechial rash on the lower extremities, arthritis of the ankles, wrists and elbows, and loose stools. He completed the amoxicillin with no worsening of symptoms. A vasculitis assessment in the Internal Medicine Clinic found a slightly elevated ANA and normal ANCAs, hepatitis B/C/HIV serologies, CH50, C3, C4, rheumatoid factor, CBC, electrolytes, coagulation, urinalysis and chest x-ray. Skin biopsy confirmed a neutrophilic small-vessel leukocytoclastic vasculitis (Figure 1). The skin rash and arthritis resolved over the next 4-6 weeks with residual hyperpigmentation and scarring. The symptoms were attributed to a possible drug allergy to amoxicillin and avoidance was recommended. Two months later, fever and pharyngitis recurred and a similar reaction occurred within 48 hours of azithromycin treatment (Figure 2). A referral was made the Adverse Drug Reaction clinic. IgE-mediated symptoms were absent. Previous treatments with penicillin were tolerated.