Controversies in surgical pathology: minimal involvement of sentinel lymph node in breast carcinoma: prevailing concepts and challenging problems

Sentinel lymph node biopsy has attained "standard of care' status in the management of breast carcinoma. However, the pathological interpretation and clinical consequences of "minimally involved' sentinel lymph nodes remain controversial. Herein, we present some of the complex and challenging pathological problems inherent in this evolving setting. Clearly, at least some of our current concepts regarding "minimally involved' sentinel lymph nodes need reappraisal.     

Characterization and physical map of choleraphage φ149 DNA

Choleraphage φ149 DNA is a linear double-stranded molecule 69 × 10⁶ Da or 104 kilobase pairs (kbp). From restriction enzyme analysis, it has been concluded that the DNA is circularly permuted. There are at least three S1 nuclease-sensitive sites along the length of the molecule. These sites represent single-strand interruptions repairable by T4 DNA ligase. A physical map of the DNA has been constructed using the restriction endonucleases BamH1 and BglII.     

Detection of chromosomes and estimation of aneuploidy in human spermatozoa using fluorescence in-situ hybridization

The development and application of fluorescence in-situ hybridization (FISH) has opened the way for comprehensive studies on numerical chromosome abnormalities in human spermatozoa. FISH can be rapidly applied to large numbers of spermatozoa and thus overcomes the major limitation of karyotyping spermatozoa after penetration of zona-free hamster oocytes. The simultaneous hybridization of two or more chromosome-specific probes to spermatozoa and subsequent detection of the bound probes using different fluorescent detection systems enables two or more chromosomes to be localized simultaneously in the same spermatozoon and provides a technique for undertaking reasonable estimates of aneuploidy. The most commonly used probes are those which bind to the centromeric region of specific chromosomes. Most studies to date have concentrated on estimating aneuploidy in spermatozoa from normospermic men, although reports are beginning to appear on aneuploidy in spermatozoa from subfertile and infertile men. Multi- probe FISH studies have generally reported disomy (hyperhaploidy) estimates of 0.05-0.2% per chromosome. There is preliminary evidence that some chromosomes such as X, Y and 21 are predisposed towards higher rates of non-disjunction during spermatogenesis. There are also suggestions of inter-donor variability in aneuploidy frequencies for specific chromosomes, although this requires confirmation in larger studies. While FISH is clearly a powerful technique that has many applications in reproductive medicine, it must also be realized that it does have limitations and the technology itself is still evolving and has yet to be fully validated on spermatozoa.      

Evaluation of two commercially available anti human immunodeficiency virus antibody ELISA Kits using clinical samples

Laboratory diagnosis is the mainstay in the diagnosis of HIV infection in an individual. The wide range of diagnostic kits available, for the detection of anti HIV antibodies, makes it mandatory that the kits are evaluated before they are made available in the market. We evaluated the performance of a Microlisa HIV kit (J. Mitra & Co.) using a panel of HIV reactive and non reactive clinical specimens. The sensitivity and specificity of the kit were found to be 100% as compared to the UBI HIV 1/2 EIA kit. The ease of testing, the assay characteristics including efficiency of the Microlisa favour its use by laboratories as a primary screen for HIV infection.     

Crosslinked chitosan: its physical properties and the effects of matrix stiffness on chondrocyte cell morphology and proliferation

Chitosan [beta(1-4)-2 amino-2-deoxy-D-glucose], the natural polyaminosaccharide derived from N-deacetylation of chitin [beta(1-4)-2 acetamide-2-deoxy-D-glucose], has been shown to possess attractive biological and cell interactive properties. Recently chitosan and chitosan analogs have also been shown to support the growth and continued function of chondrocytes. In the present study, chitosan substrates are crosslinked with a functional diepoxide (1,4 butanediol diglycidyl ether) to alter its mechanical property, and the viability and proliferation of the canine articular chondrocytes seeded on the crosslinked surface are further assayed. Of interest is the impact of substrate stiffness on the growth and proliferation of articular canine chondrocytes. Crosslinked scaffolds were also subjected to degradation by chitosanase to examine the impact of crosslinking on enzyme-assisted degradation. The hydrophilicity and compression modulus of the crosslinked surfaces were measured via contact-angle measurements and compression tests, respectively. Scanning electron microscopy (SEM) and fluorescent staining were used to observe the proliferation and morphology of chondrocyte cells on noncrosslinked and crosslinked surfaces. The crosslinked chitosan was found to be nontoxic to chondrocytes and more hydrophilic. Its compression modulus and stiffness increased, which may improve the scaffold resistance to wear and in vivo shrinkage once implanted. The increased stiffness also seemed to serve as an additional mechanical stimulus to promote chondrocyte growth and proliferation. The cell morphology on crosslinked scaffolds seen by SEM and fluorescent stain was the typical chondrocytic rounded shape. The method proposed provides a nontoxic way to increase the mechanical strength of the chitosan scaffolds.     

Sequencing of the alpha-synuclein gene in a large series of cases of familial Parkinson's disease fails to reveal any further mutations. The European Consortium on Genetic Susceptibility in Parkinson's Disease (GSPD)

A mutation in exon 4 of the human alpha-synuclein gene was reported recently in four families with autosomal dominant Parkinson's disease (PD). In order to examine whether mutations in this exon or elsewhere in the gene are common in familial PD, all seven exons of the alpha- synuclein gene were amplified by PCR from index cases of 30 European and American Caucasian kindreds affected with autosomal dominant PD. Each product was sequenced directly and examined for mutations in the open reading frame. No mutations were found in any of the samples examined. We conclude that the A53T change described in the alpha- synuclein gene is a rare cause of PD or may even be a rare variant. Mutations in the regulatory or intronic regions of the gene were not excluded by this study.      

Preliminary observations on polar body extrusion and pronuclear formation in human oocytes using time-lapse video cinematography

In this study, we have used time-lapse video cinematography to study fertilization in 50 human oocytes that had undergone intracytoplasmic sperm injection (ICSI). Time-lapse recording commenced shortly after ICSI and proceeded for 17-20 h. Oocytes were cultured in an environmental chamber which was maintained under standard culture conditions. Overall, 38 oocytes (76%) were fertilized normally, and the fertilization rate and embryo quality were not significantly different from 487 sibling oocytes cultured in a conventional incubator. Normal fertilization followed a defined course of events, although the timing of these events varied markedly between oocytes. In 35 of the 38 fertilized oocytes (92%), there were circular waves of granulation within the ooplasm which had a periodicity of 20-53 min. The sperm head decondensed during this granulation phase. The second polar body was then extruded, and this was followed by the central formation of the male pronucleus. The female pronucleus formed in the cytoplasm adjacent to the second polar body at the same time as, or slightly after, the male pronucleus, and was subsequently drawn towards the male pronucleus until the two abutted. Both pronuclei then increased in size, the nucleoli moved around within the pronuclei and some nucleoli coalesced. During pronuclear growth, the organelles contracted from the cortex towards the centre of the oocyte, leaving a clear cortical zone. The oocyte decreased in diameter from 112 to 106 microm (P < 0.0001) during the course of the observation period. The female pronucleus was significantly smaller in diameter than the male pronucleus (24.1 and 22.4 microm respectively, P = 0.008) and contained fewer nucleoli (4.2 and 7.0 respectively, P < 0.0001). After time-lapse recording, oocytes were cultured for 48 h prior to embryo transfer or cryopreservation. Embryo quality was related to fertilization events and periodicity of the cytoplasmic wave, and it was found that good quality embryos arose from oocytes that had more uniform timing from injection to pronuclear abuttal and tended to have a longer cytoplasmic wave. In conclusion, we have shown that time-lapse video cinematography is an excellent tool for studying fertilization and early embryo development, and have demonstrated that human fertilization comprises numerous complex dynamic events.      

Malassezia furfur in a case of onychomycosis: colonizer or etiologic agent?

The etiologic role of Malassezia furfur in onychomycosis is a contentious diagnostic problem because its keratinolytic ability has never been verified. This case report describes the isolation of M. furfur from the infected nails of a child clinically diagnosed with onychomycosis, and discusses the role of this organism as an etiologic agent/colonizer. The patient presented with subungual hyperkeratosis and onycholysis without associated paronychia. Budding yeast cells compatible with M. furfur were repeatedly demonstrated in KOH wet mounts of damaged nails, histopathology of hematoxylin and eosin (H&E) and periodic acid-Schiff (PAS) stained sections showed penetration of fungal elements between deeper layers of keratin, and numerous colonies of M. furfur were isolated on three consecutive occasions from nail specimens collected from different areas of hand and toenail lesions. No evidence of nail invasion by dermatophytic or nondermatophytic filamentous fungi were found by direct microscopy or culture. Microscopy and culture were negative following 12 weeks of ketoconazole treatment, which resulted in growth of healthy nail plates with normal beds. We can infer from these observations that M.furfur was an etiologic agent rather than a colonizer in the patient's nails even though direct keratinolytic character of this fungus was not demonstrated.     

Overview and update on molecular testing in non-small cell lung carcinoma utilizing endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) samples

Lung carcinoma remains one of the most frequent and aggressive human neoplasms. Fortunately, in the last decades, the increasing knowledge of the molecular mechanisms leading to cancer development has allowed the use of targeted therapies with improvement of prognosis in many patients. Clinical management has also changed after the introduction of endobronchialultrasonographic bronchoscopy that allows a conservative staging of lung tumors, avoiding the need of mediastinoscopy for lymph node staging. Lung pathologists and cytopathologists are facing the challenge of giving the more comprehensive prognostic and predictive information with ever smaller tissue or cytological samples. The aim of this review is to summarize the molecular testing for non-small cell lung carcinoma and how pathologists can contribute to the patient's outcome with a conscious management of biological samples.     

重视儿童伤害预防 落实有效干预措施

伤害是全球儿童面临的健康威胁,也是中国1~17岁儿童的首位死因。儿童伤害预防是非常具有投入产出效益的一项公共卫生措施,国内外多年的研究和实践也证明相关儿童伤害干预措施的有效性。建议加强伤害防控工作的部门分工与协调机制,确保必要的财政经费支持,以广泛实施各项有效的干预措施,保障儿童健康成长。