Breast implant-associated anaplastic large cell lymphoma in a Li-FRAUMENI patient: a case report.

BACKGROUND: Breast implant-associated anaplastic large cell lymphoma (BIA-ALCL) is a rare malignancy, recently recognized as a provisional entity by the World Health Organization. Although increasing data have been published on this entity in recent years, a great number of patients and health professionals remain unaware of this diagnosis. CASE PRESENTATION: We herein report the case of a 56-year-old female with Li-FRAUMENI syndrome who presented with late right-sided recurrent breast swelling after prophylactic adenomastectomy with implant reconstruction. Imaging scans revealed an heterogeneous mass adjacent to the implant fibrous capsule. A biopsy of the lesion rendered the diagnosis of a BIA-ALCL. CONCLUSIONS: This case presents similarities with previous reports, but also some particularities, which should be stressed in order to make the diagnosis the earliest possible. The most distinct feature is that this is the second report of BIA-ALCL arising in the setting of Li-FRAUMENI syndrome.

Cathepsin K induces platelet dysfunction and affects cell signaling in breast cancer - molecularly distinct behavior of cathepsin K in breast cancer.

BACKGROUND: Breast cancer comprises clinically and molecularly distinct tumor subgroups that differ in cell histology and biology and show divergent clinical phenotypes that impede phase III trials, such as those utilizing cathepsin K inhibitors. Here we correlate the epithelial-mesenchymal-like transition breast cancer cells and cathepsin K secretion with activation and aggregation of platelets. Cathepsin K is up-regulated in cancer cells that proteolyze extracellular matrix and contributes to invasiveness. Although proteolytically activated receptors (PARs) are activated by proteases, the direct interaction of cysteine cathepsins with PARs is poorly understood. In human platelets, PAR-1 and -4 are highly expressed, but PAR-3 shows low expression and unclear functions. METHODS: Platelet aggregation was monitored by measuring changes in turbidity. Platelets were immunoblotted with anti-phospho and total p38, Src-Tyr-416, FAK-Tyr-397, and TGFß monoclonal antibody. Activation was measured in a flow cytometer and calcium mobilization in a confocal microscope. Mammary epithelial cells were prepared from the primary breast cancer samples of 15 women with Luminal-B subtype to produce primary cells. RESULTS: We demonstrate that platelets are aggregated by cathepsin K in a dose-dependent manner, but not by other cysteine cathepsins. PARs-3 and -4 were confirmed as the cathepsin K target by immunodetection and specific antagonists using a fibroblast cell line derived from PARs deficient mice. Moreover, through co-culture experiments, we show that platelets activated by cathepsin K mediated the up-regulation of SHH, PTHrP, OPN, and TGFß in epithelial-mesenchymal-like cells from patients with Luminal B breast cancer. CONCLUSIONS: Cathepsin K induces platelet dysfunction and affects signaling in breast cancer cells.

Angiomirs expression profiling in diffuse large B-Cell lymphoma.

Despite advances in treatment, 30% of diffuse large B-cell lymphoma (DLBCL) cases are refractory or relapse after chemoimmunotherapy. Currently, the relationship between angiogenesis and angiomiRs in DLBCL is unknown. We classified 84 DLBCL cases according to stromal signatures and evaluated the expression of pro- and antiangiomiRs in paraffin embedded tissues of DLBCL and correlated them with microvascular density (MVD). 40% of cases were classified as stromal-1, 50% as stromal-2 and 10% were not classified. We observed increased expression of proangiomiRs Let-7f, miR-17, miR-18a, miR-19b, miR-126, miR-130a, miR-210, miR-296 and miR-378 in 14%, 57%, 30%, 45%, 12%, 12%, 56%, 58% and 48% of the cases, respectively. Among antiangiomiRs we found decreased expression of miR-16, miR-20b, miR-92a, miR-221 and miR-328 in, respectively, 27%, 71%, 2%, 44% and 11%. We found association between increased expression of proangiomiRs miR-126 and miR-130a and antiangiomiR miR-328 and the subtype non-GCB. We found higher levels of the antiangiomiRs miR-16, miR-221 and miR-328 in patients with low MVD and stromal-1 signature. IPI and CD34 confirmed independent impact on survival of the study group. None of the above angiomiRs showed significance as biomarker in an independent serum samples cohort of patients and controls. In conclusion, we confirmed association between antiangiomiRs miR-16, miR-221 and miR-328 and stromal-1 signature. Four angiomiRs emerged as potential therapeutic targets: proangiomiRs miR-17, miR-210 and miR-296 and antiangiomiR miR-20b. Although the four microRNAs seem to be important in DLBCL pathogenesis, they were not predictive of DLBCL onset or relapse in the serum independent cohort.

Pathologic and imunohistochemical characterization of tumoral inflammatory cell infiltrate in invasive penile squamous cell carcinomas: Fox-P3 expression is an independent predictor of recurrence.

Penile carcinomas (PeCa) are relatively rare, but devastating neoplasms, more frequent among people of underprivileged socioeconomic status. There is mounting evidence that immune cells may trigger various mechanisms that enhance tumor growth and metastasis, but no data on the peritumoral inflammation is available for PeCa. The objectives of the present study are to evaluate the immunohistomorphology of tumoral inflammation in PeCa, and to correlate it with clinicopathological parameters, which could contribute to the prognostic evaluation. One hundred and twenty-two patients with the diagnosis of usual-type squamous cell penile carcinoma were included. Paraffin-embedded tissue was submitted to immunohistochemical evaluation of p16 protein, CD3, CD4, CD8, CD20, CD68, CD138, granzyme B, and Fox-P3. The Fisher's exact test was employed for comparison between histological variables and parameters, and the Kaplan-Meier method for the analysis of survival. Improved 5-year overall survival was significantly associated to age ≤60 years, stage I + II, tumor size T1 + T2, lymph node status N0, and absent perineural invasion. In a multivariate analysis age ≥60 years, presence of lymph node metastasis, urethral invasion, and high histologic grade retained a significantly more unfavorable outcome. Improved 5-year failure free survival was associated to stage of the disease I + II, lymph node status N0, absence of perineural, vascular, and urethral invasion, and Fox-P3 expression. In a multivariate analysis, presence of lymph node metastasis, perineural and vascular invasion, and of Fox-P3-positive lymphocytes together with low inflammatory infiltrate retained a significantly more unfavorable outcome. These results support the prognostic value of determining the levels of Fox-P3-positive lymphocytes by immunohistochemistry in PeCa, as this parameter adds value to the traditional clinicopathological features.

Biobanking practice: RNA storage at low concentration affects integrity.

A critical issue in defining protocols for biobanking practices is the preservation of total RNA for assessing the whole transcriptome and ensuring that it can be utilized in clinically oriented studies. Storage conditions, such as temperature and the length of time that tissues and purified RNA stay frozen, may directly impact RNA preservation. In this study, we evaluated a) the quality of RNA (as measured by RNA Integrity Number) purified from head and neck tumor tissues stored at -140°C for distinct time intervals of up to 7 years, and b) the quality of their respective RNAs stored for 4 years at -80°C when diluted at either 250 ng/µL or 25 ng/µL, with repeated freezing and thawing. Additionally, we generated a profile of the RNA collection of human tumors from different body sites stored at the AC Camargo Biobank. Our results showed no significant change in RIN values according to length of storage at -140°C. With respect to RNA aliquots stored at -80°C, RNA integrity at 250 ng/µL was preserved, while statistically significant degradation was observed at 25 ng/µL after only 8 months of storage. The RNA collection from most of the human tumors stored at the AC Camargo Biobank exhibited high quality, with average RIN around seven. However, ovary and stomach samples had the greatest RNA degradation. Taken together, the results show that both the temperature of preservation and the concentration of RNA should be strictly controlled by the biobank staff involved in macromolecule purification. Moreover, the RNAs from our biobank can be useful for the most demanding methods of gene expression analysis by virtue of adherence to optimal standard operating procedures for both tissue and macromolecule laboratories.

Identification of upregulated genes in oral squamous cell carcinomas.

BACKGROUND: Oral cancer is the most common subset of head and neck squamous cell carcinomas (HNSCC). These tumors often have an aggressive clinical outcome hallmarked by a propensity for local invasion and regional nodal metastasis. Upregulated genes could be useful as markers for diagnosis, prognosis, and as new drug targets for these tumors. METHODS: To identify upregulated genes in oral squamous cell carcinomas (OSSCs), we examined the ORESTES public database and used a quantitative reverse transcription-polymerase chain reaction (qRT-PCR) approach to determine the expression level of selected genes in tumor samples. RESULTS AND CONCLUSIONS: The ORESTES data mining analysis indicated 40 upregulated genes in HNSCC. Nine of these candidate genes were selected for further qRT-PCR validation and 3 of them (ALDOA, AHSA1, and POLQ) were frequently found upregulated in OSCC samples, which may indicate an association of these genes with the carcinogenesis process in this tumor site and they can constitute potential new targets for therapy.

Rural-origin health science students at South African universities.

BACKGROUND: Rural areas in all countries suffer from a shortage of health care professionals. In South Africa, the shortage is particularly marked; some rural areas have a doctor-to-population ratio of 5.5:100 000. Similar patterns apply to other health professionals. Increasing the proportion of rural-origin students in faculties of health sciences has been shown to be one way of addressing such shortages, as the students are more likely to work in rural areas after graduating. OBJECTIVE: To determine the proportion of rural-origin students at all medical schools in South Africa. DESIGN: A retrospective descriptive study was conducted in 2003. Lists of undergraduate students admitted from 1999 to 2002 for medicine, dentistry, physiotherapy and occupational therapy were obtained from 9 health science faculties. Origins of students were classified as city, town and rural by means of postal codes. The proportion of rural-origin students was determined and compared with the percentage of rural people in South Africa (46.3%). RESULTS: . Of the 7 358 students, 4 341 (59%) were from cities, 1 107 (15%) from towns and 1 910 (26%) from rural areas. The proportion of rural-origin students in the different courses nationally were: medicine--27.4%, physiotherapy--22.4%, occupational therapy--26.7%, and dentistry--24.8%. CONCLUSION: The proportion of rural-origin students in South Africa was considerably lower than the national rural population ratio. Strategies are needed to increase the number of rural-origin students in universities via preferential admission to alleviate the shortage of health professionals in rural areas.

Key issues in clinic functioning - a case study of two clinics.

OBJECTIVE: The aim of this research was to understand key issues in the functioning of two different primary care clinics serving the same community, in order to learn more about clinic management. DESIGN: An in-depth case study was conducted. A range of qualitative information was collected at both clinics. Data collected in the two clinics were compared, to gain an understanding of the important issues. SETTING: Data were collected in a government and an NGO clinic in North West province. SUBJECTS: This report presents the findings from patient and staff satisfaction surveys and in-depth individual interviews with senior staff. RESULTS: Key findings included the following: (i) there are attitudinal differences between the staff at the two clinics; (ii) the patients appreciate the services of both clinics, though they view them differently; (iii) clinic A provides a wider range of services to more people more often; (iv) clinic B presents a picture of quality of care, related to the environment and approach of staff; (v) waiting time is not as important as how patients are treated; (vi) medications are a crucial factor, in the minds of staff and patients; and (vii) a supportive, empowering organisational culture is needed to encourage staff to deliver better care to their patients. The management of the clinic is part of this culture. CONCLUSIONS: This research provides lessons regarding key issues in clinic functioning which can make a major difference to the way services are experienced. A respectful and caring approach to patients, and an organisational culture which supports and enables staff, can achieve much of this without any additional resources.

Influences on the choice of health professionals to practice in rural areas.

BACKGROUND: Training health care professionals (HCPs) to work in rural areas is a challenge for educationalists. This study aimed to understand how HCPs choose to work in rural areas and how education influences this. METHODS: Qualitative individual interviews were conducted with 15 HCPs working in rural areas in SA. RESULTS: Themes identified included personal, facilitating, contextual, staying and reinforcing factors. Personal attributes of the HCPs, namely rural origin and/or their value system, determine consideration of rural practice. The decision to 'go rural' is facilitated by exposure to rural practice during training, an understanding of rural needs and exposure to rural role models. Once practising in a rural area, the context and nature of work and the environment influence the decision to remain, supported by the role of family and friends, ongoing training and development, and the style of health service management. Personal motivation is reinforced by a positive relationship with the community, and by being an advocate and role model for the local community. Educational factors were often felt to work against the decision to practise in rural areas. DISCUSSION: The results show the complexity of the interaction between a large number of factors working together to make HCPs choose to go and stay in rural areas. Factors other than educational ones seem more important. A comprehensive approach is needed to attract and retain HCPs in rural areas. Issues for educationalists to address include helping rural-origin students to connect with their own values and communities.

The involvement of private general practitioners in visiting primary healthcare clinics

Background: The primary healthcare system was adopted as the vehicle of healthcare delivery and a means of reaching the larger part of the population in South Africa in 1994. One of the strategies employed in providing a comprehensive service is the incorporation of visits to clinics by doctors in support of other members of the primary healthcare team; particularly nurses. A successful collaboration at this level brings benefit to everyone involved; particularly patients. Clear expectations and a confusion of roles leads to lack of teamwork; thus it is important to have clearly established models for such involvement. Doctors working in district hospitals mostly visit clinics; but their workload; staff shortages and transport often interfere with these visits. As a form of private-public partnership; local GPs are sometimes contracted to visit the clinics. Very little is known about this practice and problems are reported; including the perception that GPs do not spend as much time in the clinics as they are paid for10.Understanding the practice better may provide answers on how to improve the quality of primary care in the district health system. The aim of this study was to describe the experiences of local GPs visiting public clinics regularly over a long period of time. Methods A case study was undertaken in the Odi district of the North West Province in three primary care clinics visited by GPs. The experiences of the doctors; clinic nurses; district managers and patients regarding the GP's visits were elicited through in-depth interviews. Details of the visits with regard to patient numbers; lengths of the visits; remuneration and preferences were also sought. The data were analysed using different methods to highlight important themes. Results: The visits by the GPs to the clinics were viewed as beneficial by the patients and clinic staff. The GPs were often preferred to government doctors because of their skills; patience and availability. The visits were also seen as a gesture of patriotism by the GPs. There were constraints; such as a shortage of medicines and equipment; which reduce the success of these visits. Conclusion: The involvement of GPs in primary care clinics is beneficial and desirable. It enhances equity in terms of access to services. Addressing the constraints can optimise the public-private partnership at this level

The Involvement of Private General Practitioners in Visiting Primary Healthcare Clinics

Background: The primary healthcare system was adopted as the vehicle of healthcare delivery and a means of reaching the larger part of the population in South Africa in 1994. One of the strategies employed in providing a comprehensive service is the incorporation of visits to clinics by doctors in support of other members of the primary healthcare team; particularly nurses. A successful collaboration at this level brings benefit to everyone involved; particularly patients. Clear expectations and a confusion of roles leads to lack of teamwork; thus it is important to have clearly established models for such involvement. Doctors working in district hospitals mostly visit clinics; but their workload; staff shortages and transport often interfere with these visits. As a form of private-public partnership; local GPs are sometimes contracted to visit the clinics. Very little is known about this practice and problems are reported; including the perception that GPs do not spend as much time in the clinics as they are paid for10.Understanding the practice better may provide answers on how to improve the quality of primary care in the district health system. The aim of this study was to describe the experiences of local GPs visiting public clinics regularly over a long period of time.Methods: A case study was undertaken in the Odi district of the North West Province in three primary care clinics visited by GPs. The experiences of the doctors; clinic nurses; district managers and patients regarding the GP's visits were elicited through in-depth interviews. Details of the visits with regard to patient numbers; lengths of the visits; remuneration and preferences were also sought. The data were analysed using different methods to highlight important themes.Results: The visits by the GPs to the clinics were viewed as beneficial by the patients and clinic staff. The GPs were often preferred to government doctors because of their skills; patience and availability. The visits were also seen as a gesture of patriotism by the GPs. There were constraints; such as a shortage of medicines and equipment; which reduce the success of these visits.Conclusion: The involvement of GPs in primary care clinics is beneficial and desirable. It enhances equity in terms of access to services. Addressing the constraints can optimise the public-private partnership at this level

The neonatal resuscitation training project in rural South Africa.

A paediatrician trainer from Australia (JT) spent 3 months in South Africa to assist with the development of neonatal resuscitation training in rural areas, particularly in district hospitals. The project was initiated by the Rural Health Unit at the University of the Witwatersrand and coordinated through the Family Medicine Education Consortium (FaMEC). The Rural Workforce Agency of Victoria together with General Practice and Primary Health Care Northern Territory covered the salary and international travel costs of the trainer, while local costs were funded by provincial departments of health, participants and a Belgian funded FaMEC project. The trainer developed an appropriate one-day skills training course in neonatal resuscitation (NNR), using the South African Paediatric Association Manual of Resuscitation of the Newborn as pre-reading, and a course to train trainers in neonatal resuscitation. From July to October 2004 he moved around the country running the neonatal resuscitation course, and, more importantly, training and accrediting trainers to run their own courses on an ongoing basis. The neonatal resuscitation course involved pre- and post-course multiple-choice question tests to assess knowledge and application, and, later, pre- and post-course skills tests to assess competence. A total of 415 people, including 215 nurses and 192 doctors, attended the neonatal resuscitation courses in 28 different sites in eight provinces. In addition, 97 trainers were trained, in nine sites. The participants rated the course highly. Pre- and post-course tests showed a high level of learning and improved confidence. The logistical arrangements, through the departments of family medicine, worked well, but the programme was very demanding of the trainer. Lessons and experiences were not shared between provinces, leading to repetition of some problems. A clear issue around the country was a lack of adequate equipment in hospitals for neonatal resuscitation, which needs to be addressed by health authorities. A process of ongoing training has been established, with provincial coordinators taking responsibility for standards and the roll-out of training. A formal evaluation of the project is planned. The project serves as a model for skills training in rural areas in South Africa, and for collaboration between organisations. A number of specific recommendations are made for the future of this NNR training project, which offer lessons for similar programmes.

Expressão imuno - histoquímica de c - erb - B2 e p53 em carcinomas gástricos / Imunohistochemical expression of c - erb - B2 and p53 in gastric carcinomas

INTRODUÇAO: Em nosso meio, os carcinomas gástricos ainda são neoplasias bastante freqüentes e responsáveis por altas taxas de mortalidade. Recentemente, têm-se demonstrado a expressão de p53 e a amplificacão do gene c-erb-B2 nos carcinomas gástricos. A relevância e o significado biológico destas alteracões ainda não foram totalmente estabelecidos. OBJETIVO: Estudar as expressões imuno-histoquímicas de p53 e c-erb-B2 em 482 casos de carcinomas gástricos. MATERIAL E MÉTODOS: Foram construídos três blocos de tissue microarray (TMA) utilizando-se duplicatas de 482 casos de carcinomas gástricos. Os cortes foram corados por hematoxilina e eosina (HE), tendo sido feita pesquisa para p53 e c-erb-B2. Foram considerados positivos para p53 os casos com marcacão nuclear em mais de 10 por cento das células tumorais. Para o c-erb-B2 foram considerados positivos os casos com marcacão de membrana completa em mais de 10 por cento das células tumorais. RESULTADOS: A expressão de p53 e c-erb-B2 foi observada em 30 por cento e 12 por cento dos casos, respectivamente. Em relacão aos tipos histológicos observou-se correlacão entre os carcinomas do tipo intestinal e a expressão de c-erb-B2 (p < 0,001). A expressão de p53 foi mais freqüente nos carcinomas com mais de 5cm de diâmetro (p = 0,036). Não foram observadas alteracões nas curvas de sobrevida dos pacientes em relacão às expressões desses marcadores. CONCLUSAO: Em nosso meio, carcinomas gástricos do tipo intestinal são mais freqüentemente positivos para c-erb-B2 nos tipos intestinais do que nos difusos. A expressão de p53 está associada ao tamanho tumoral. A técnica do TMA é válida e eficiente para o estudo de marcadores imuno-histoquímicos, com forte correlacão com os cortes tradicionais de representacão do tumor.

Overexpression of Fos-related antigen-1 in head and neck squamous cell carcinoma.

The activating protein-1 (AP-1) family of transcription factors has been implicated in the control of proliferation and differentiation of keratinocytes, but its role in malignant transformation is not clear. The aim of this study is to assess the pattern of mRNA expression of jun-fos AP-1 family members in 45 samples of head and neck squamous cell carcinomas (HNSCC) and matched adjacent mucosa by means of Northern blot analysis. Transcripts of all family members were identified, except for JunB that was detected only by means of reverse transcription polymerase chain reaction. Neither c-Fos nor JunD or FosB mRNA differed between tumours and normal tissues. We observed a strong Fos-related antigen-1 (Fra-1) and Fra-2 expression, but only Fra-1 mRNA densitometric values were higher in tumour, compared to normal adjacent mucosa (t-test, P = 0.006). A direct relationship between the positive expression of Fra-1 mRNA, above tumour median, was associated with the presence of compromised lymph nodes (Fischer exact test, P = 0.006). In addition, Fra-1 protein staining was assessed in a collection of 180 tumours and 29 histologically normal samples adjacent to tumours in a tissue array. Weak reactivity, restricted to the basal cell layer, was detected in 79% of tumour adjacent normal tissues, opposed to the intense reactivity of cancer tissues. In the subgroup of oral cancers, we have observed a shift in Fra-1 immunoreactivity, as long as the number of patients in each category, cytoplasmic or nuclear/cytoplasmic staining, was analysed (Fischer exact test, P = 0.0005). Thus, Fra-1 gene induction and accumulation of Fra-1 protein may contribute to the neoplastic phenotype in HNSCC.