Geospatial analysis of Plasmodium falciparum serological indicators: school versus community sampling in a low-transmission malaria setting.

BACKGROUND: Due to low numbers of active infections and persons presenting to health facilities for malaria treatment, case-based surveillance is inefficient for understanding the remaining disease burden in low malaria transmission settings. Serological data through the detection of IgG antibodies from previous malaria parasite exposure can fill this gap by providing a nuanced picture of where sustained transmission remains. Study enrollment at sites of gathering provides a potential approach to spatially estimate malaria exposure and could preclude the need for more intensive community-based sampling. METHODS: This study compared spatial estimates of malaria exposure from cross-sectional school- and community-based sampling in Haiti. A total of 52,405 blood samples were collected from 2012 to 2017. Multiplex bead assays (MBAs) tested IgG against P. falciparum liver stage antigen-1 (LSA-1), apical membrane antigen 1 (AMA1), and merozoite surface protein 1 (MSP1). Predictive geospatial models of seropositivity adjusted for environmental covariates, and results were compared using correlations by coordinate points and communes across Haiti. RESULTS: Consistent directional associations were observed between seroprevalence and environmental covariates for elevation (negative), air temperature (negative), and travel time to urban centers (positive). Spearman's rank correlation for predicted seroprevalence at coordinate points was lowest for LSA-1 (ρ = 0.10, 95% CI: 0.09-0.11), but improved for AMA1 (ρ = 0.36, 95% CI: 0.35-0.37) and MSP1 (ρ = 0.48, 95% CI: 0.47-0.49). CONCLUSIONS: In settings approaching P. falciparum elimination, case-based prevalence data does not provide a resolution of ongoing malaria transmission in the population. Immunogenic antigen targets (e.g., AMA1, MSP1) that give higher population rates of seropositivity provide moderate correlation to gold standard community sampling designs and are a feasible approach to discern foci of residual P. falciparum transmission in an area.

Nurse-led dexmedetomidine sedation for magnetic resonance imaging in children: a 6-year quality improvement project.

We aimed to safely introduce dexmedetomidine into a nurse-led sedation service for magnetic resonance imaging in children. Secondary aims were to increase the number of children eligible for sedation and to increase the actual number of children having sedation performed by our nurse sedation team. We analysed 1768 consecutive intravenous and 219 intranasal dexmedetomidine sedation episodes in infants, children and adolescents having magnetic resonance imaging scans between March 2016 and March 2022. The overall sedation success rate was 98.4%, with a 98.9% success rate for intravenous dexmedetomidine and a 95.0% success rate for intranasal dexmedetomidine. The incidence of scan interruption during intravenous and intranasal dexmedetomidine sedation was 8.8% and 21.9%, respectively. We conclude that paediatric sedation with dexmedetomidine for magnetic resonance scanning is safe and successful.

Development of a new peptide-bead coupling method for an all peptide-based Luminex multiplexing assay for detection of Plasmodium falciparum antibody responses.

Using a recombinant protein antigen for antibody testing shows a sum of antibody responses to multiple different immune epitopes existing in the protein antigen. In contrast, the antibody testing to an immunogenic peptide epitope reflects a singular antibody response to the individual peptide epitope. Therefore, using a panel of peptide epitopes provides an advantage for profiling multiple singular antibody responses with potential to estimate recent malaria exposure in human infections. However, transitioning from malaria immune epitope peptide-based ELISA to an all peptide bead-based multiplex Luminex assay presents some challenges including variation in the ability of different peptides to bind beads. The aim of this study was to develop a peptide coupling method while demonstrating the utility of these peptide epitopes from multiple stage antigens of Plasmodium falciparum for measuring antibodies. Successful coupling of peptide epitopes to beads followed three steps: 1) development of a peptide tag appended to the C-terminus of each peptide epitope consisting of beta-alanine-lysine (x 4)--cysteine, 2) bead modification with a high concentration of adipic acid dihydrazide, and 3) use of the peptide epitope as a blocker in place of the traditional choice, bovine serum albumin (BSA). This new method was used to couple 12 peptide epitopes from multiple stage specific antigens of P. falciparum, 1 Anopheles mosquito salivary gland peptide, and 1 Epstein-Barr virus peptide as an assay control. The new method was applied to testing of IgG in pooled samples from 30 individuals with previously repeated malaria exposure in western Kenya and IgM and IgG in samples from 37 U.S. travelers with recent exposure to malaria. The new peptide-bead coupling method and subsequent multiplex Luminex assay showed reliable detection of IgG to all 14 peptides in Kenyan samples. Among 37 samples from U.S. travelers recently diagnosed with malaria, IgM and IgG to the peptide epitopes were detected with high sensitivity and variation. Overall, the U.S. travelers had a much lower positivity rates of IgM than IgG to different peptide epitopes, ranging from a high of 62.2% positive for one epitope to a low of only 5.4% positive for another epitope. In contrast, the travelers had IgG positive rates from 97.3% to 91.9% to various peptide epitopes. Based on the different distribution in IgM and IgG positivity to overall number of peptide epitopes and to the number of pre-erythrocytic, erythrocytic, gametocytic, and salivary stage epitopes at the individual level, four distinct patterns of IgM and IgG responses among the 37 samples from US travelers were observed. Independent peptide-bead coupling and antibody level readout between two different instruments also showed comparable results. Overall, this new coupling method resolves the peptide-bead coupling challenge, is reproducible, and can be applied to any other immunogenic peptide epitopes. The resulting all peptide bead-based multiplex Luminex assay can be expanded to include other peptide epitopes of P. falciparum, different malaria species, or other diseases for surveillance, either in US travelers or endemic areas.

Surgical outcomes after radioactive 125I seed versus hookwire localization of non-palpable breast cancer: a multicentre randomized clinical trial.

BACKGROUND: Previous studies have suggested improved efficiency and patient outcomes with 125I seed compared with hookwire localization (HWL) in breast-conserving surgery, but high-level evidence of superior surgical outcomes is lacking. The aim of this multicentre pragmatic RCT was to compare re-excision and positive margin rates after localization using 125I seed or hookwire in women with non-palpable breast cancer. METHODS: Between September 2013 and March 2018, women with non-palpable breast cancer eligible for breast-conserving surgery were assigned randomly to preoperative localization using 125I seeds or hookwires. Randomization was stratified by lesion type (pure ductal carcinoma in situ (DCIS) or other) and study site. Primary endpoints were rates of re-excision and margin positivity. Secondary endpoints were resection volumes and weights. RESULTS: A total of 690 women were randomized at eight sites; 659 women remained after withdrawal (125I seed, 327; HWL, 332). Mean age was 60.3 years in the 125I seed group and 60.7 years in the HWL group, with no difference between the groups in preoperative lesion size (mean 13.2 mm). Lesions were pure DCIS in 25.9 per cent. The most common radiological lesion types were masses (46.9 per cent) and calcifications (28.2 per cent). The localization modality was ultrasonography in 65.5 per cent and mammography in 33.7 per cent. The re-excision rate after 125I seed localization was significantly lower than for HWL (13.9 versus 18.9 per cent respectively; P = 0.019). There were no significant differences in positive margin rates, or in specimen weights and volumes. CONCLUSION: Re-excision rates after breast-conserving surgery were significantly lower after 125I seed localization compared with HWL. Registration number: ACTRN12613000655741 (http://www.ANZCTR.org.au/).

A text message intervention to support women's physical and mental health after breast cancer treatments (EMPOWER-SMS): a randomised controlled trial protocol.

BACKGROUND: Breast cancer is the most common cancer diagnosed in women worldwide. In developed countries, 80-90% of women will survive five years after diagnosis but the transition from hospital-based care to health self-management and self-efficacy can be difficult. Text messaging programs offer a simple and proven way to provide support to people with chronic diseases. This study aims to test the effectiveness of a text message support program at improving women's health self-efficacy, and physical and mental health outcomes after breast cancer treatments compared to usual care at 6-months and to understand the barriers and enablers to widespread implementation. METHODS: Single-blind randomised control trial (RCT; N = 160) comparing a text message support intervention to usual care in women with breast cancer (recruited from a large tertiary referral hospital in Sydney, Australia). The intervention group will receive a six-month text message support program, which consists of semi-personalised, supportive, lifestyle-focused text messages (4 messages/week) in addition to usual care. The control group will receive usual care without the text message program. Outcomes will be assessed at 6-months. The primary outcome is change in self-efficacy for managing chronic disease. Secondary outcomes include change in clinical outcomes (body mass index), lifestyle outcomes (physical activity levels, dietary behaviours), mood (depression and anxiety scales), quality of life, satisfaction with, and usefulness of the intervention. Analyses will be performed on the principle of intention-to-treat to examine differences between intervention and control groups. DISCUSSION: This study will test if a scalable and cost-effective text-messaging intervention is effective at improving women's health self-efficacy, as well as physical and mental health outcomes. Moreover, this study will provide essential preliminary data to bolster a large multicentre RCT to helpsupport breast cancer survivors throughout recovery and beyond. TRIAL REGISTRATION: Australia New Zealand Clinical Trials Registry (ANZCTR) number ACTRN12618002020268 , 17 December 2018.

Factors associated with romantic relationship formation difficulties in women with breast cancer.

OBJECTIVE: Many un-partnered women report difficulty in forming romantic relationships after breast cancer, characterized by high dating-related anxiety and low perceived interpersonal competence. This study examined the relationship between poor body image (appearance investment and body dissatisfaction) and self-compassion, and women's ability to form romantic relationships post-breast cancer. METHODS: Women (N = 152) diagnosed with breast cancer, who were either un-partnered and expressed interest in romantic dating, or who had commenced a relationship post-diagnosis, completed an online survey. Assessments included the Interpersonal Competence Questionnaire, Dating Anxiety Scale, Self-compassion Scale, Appearance Schemas Inventory-Revised, Body Image Scale, and Experiences in Close Relationships Scale. Multiple regression analyses assessed the relationships between these variables. RESULTS: Partnered and un-partnered women differed in levels of dating anxiety, interpersonal competence, anxious attachment, and the self-evaluative salience facet of appearance investment. Analyses revealed a significant model for dating anxiety, with high self-evaluative salience, body image dissatisfaction, and attachment avoidance independently associated with this outcome. The model for interpersonal competence was also significant, with low attachment avoidance and high self-compassion independently associated with this outcome. CONCLUSIONS: Un-partnered women who place high importance on appearance for their self-worth and who report poor body image and low self-compassion are at risk of experiencing difficulties in forming new romantic relationships after breast cancer. Future interventions should target these variables to facilitate romantic dating during cancer survivorship.

Patient reported outcomes of autologous fat grafting after breast cancer surgery.

INTRODUCTION: Autologous fat grafting (AFG) can be used as an adjunct in breast cancer surgery to improve contour defects. Few previous studies have assessed patient reported outcomes (PROs) for AFG. This study analysed AFG use and assessed PROs in terms of physical and psychosocial well-being. MATERIALS AND METHODS: All patients undergoing AFG were identified from a prospective database and asked to complete the validated BREAST-Q questionnaire and a tool to assess patient-perceived change after AFG (5-point Likert-type scale). Descriptive statistics were computed for all BREAST-Q and perceived change subscales. Independent sample t-tests were conducted to compare scores on each of the BREAST-Q and perceived change subscales by type of breast cancer surgery and radiotherapy status. RESULTS: 156 AFG sessions were performed over 4 years on 119 breasts in 88 patients. Fifty-seven patients received AFG after reconstruction and 19 after breast conserving surgery. Forty-six patients (52%) completed the questionnaire. BREAST-Q scores (out of 100) and patient-perceived change after AFG (out of 5) were respectively: 54 and 4.0 for Breast satisfaction, 69 and 3.3 for Physical well-being and 60 and 3.6 for Psychosocial well-being. Radiotherapy status and type of surgery made little difference. Number of AFG procedures positively correlated with perceived improvement. DISCUSSION: Autologous fat grafting was associated with improved patient satisfaction despite small volumes transferred. BREAST-Q scores were comparable with previously published series on reconstructive breast surgery. Perceived change after AFG was no different in patients receiving radiotherapy.

Role of Water Sorption in Tablet Crushing Strength, Disintegration, and Dissolution.

Drugs formulated as tablets are subjected to accelerated stability conditions with the goal of identifying a stable formulation that will exhibit a sufficiently long shelf life. Water sorption at a condition such as 40°C/75% RH can result in significant changes in tablet properties such as a decrease in dissolution rate, the cause of which may be difficult to interpret, given the complex nature of ingredients and their interactions in a tablet. In this research, three drugs, displaying a wide range of physicochemical properties, were formulated with commonly used diluents, disintegrants, and binders, using a design of experiments approach. The tablets were stored at accelerated conditions and assessed for content, dissolution, disintegration, and crushing strength, as well as other properties. The research demonstrated many water-induced effects in tablet properties. Due to the experimental design approach that revealed many interactions, it was possible to interpret all of the changes observed in tablet crushing strength, disintegration, and dissolution for the drugs using a common set of physical principles. Specifically, the relevant factors considered were (1) mechanical properties of materials, (2) water sorption surface effects in surface diffusion and capillary condensation, (3) water sorption bulk effects for amorphous materials such as viscous flow/spreading, and (4) water-induced stress on interparticle bonding arising from volume expansion. These physical principles enable a comprehensive interpretation of the complex changes observed in tablet properties, which should be valuable in the design of tablet formulations that will be stable to accelerated storage conditions.

Body image and psychological distress in nipple-sparing mastectomy: the roles of self-compassion and appearance investment.

OBJECTIVE: Women with breast cancer face threats to body image following surgery. Nipple-sparing mastectomy with immediate breast reconstruction (NSM + IBR) may minimise body image disturbance as this preserves the woman's skin and areola complex. We assessed levels of body image disturbance and psychological distress in women undergoing NSM + IBR. To further understand the body image-distress relationship, we investigated the potential moderating effect of self-compassion and appearance investment on this relationship. METHODS: Women diagnosed with breast cancer (N = 75) who had undergone NSM + IBR completed online questionnaires including the Body Image Scale, general (Depression, Anxiety and Stress Scales) and cancer-specific (Impact of Event Scale) psychological distress and Self-Compassion Scale and Appearance Schemas Inventory - Revised. RESULTS: Mean general and cancer-specific psychological distress scores were within normal ranges, and body image disturbance was moderately low. Body image was positively correlated with depression, stress, Impact of Event Scale scores and appearance investment and negatively correlated with self-compassion. MANCOVA analyses indicated a significant moderating effect of self-compassion and appearance investment on the body image disturbance-distress relationship (for depression, stress and intrusion), such that participants with high self-compassion and low appearance investment experienced lower distress than individuals with low self-compassion and high appearance investment. CONCLUSIONS: Moderately low levels of psychological distress and body image disturbance suggest NSM + IBR may minimise adverse psychological impacts of mastectomy. Increased body image disturbance was associated with psychological distress and moderated by self-compassion and appearance investment, suggesting a potential role for these characteristics as the focus of psychological interventions to minimise the negative impacts of mastectomy. Copyright © 2016 John Wiley & Sons, Ltd.

Tissue biomarkers of breast cancer and their association with conventional pathologic features.

BACKGROUND: Tissue protein expression profiling has the potential to detect new biomarkers to improve breast cancer (BC) diagnosis, staging, and prognostication. This study aimed to identify tissue proteins that differentiate breast cancer tissue from healthy breast tissue using protein chip mass spectrometry and to examine associations with conventional pathological features. METHODS: To develop a training model, 82 BC and 82 adjacent unaffected tissue (AT) samples were analysed on cation-exchange protein chips by time-of-flight mass spectrometry. For validation, 89 independent BC and AT sample pairs were analysed. RESULTS: From the protein peaks that were differentially expressed between BC and AT by univariate analysis, binary logistic regression yielded two peaks that together classified BC and AT with a ROC area under the curve of 0.92. Two proteins, ubiquitin and S100P (in a novel truncated form), were identified by liquid chromatography/tandem mass spectrometry and validated by immunoblotting and reactive-surface protein chip immunocapture. The combined marker panel was positively associated with high histologic grade, larger tumour size, lymphovascular invasion, ER and PR positivity, and HER2 overexpression, suggesting that it may be associated with a HER2-enriched molecular subtype of breast cancer. CONCLUSION: This independently validated protein panel may be valuable in the classification and prognostication of breast cancer patients.

SU-E-T-415: Evaluation of the Effect of the Dose Buildup Region for Flattening Filter Free Beams in IMRT Plans of Lung and Sinuses.

PURPOSE: The surface and buildup dose is known to be greater for flattening-filter-free (FFF) beams as compared to flattened beams. We compare the effect of this difference in the dose buildup region between the two beam modes, in the context of IMRT plans for the lung and sinuses. METHODS: IMRT plans are delivered using both the standard flattened beam mode (6X) and flattening-filter-free mode (6XFFF). We compare the dosimetric differences of these two modes, for IMRT treatments for the lung and sinuses. Radiochromic films and parallel-plate ionization chamber measurements were taken, and evaluated for the following field sizes: 2×2 cm2 , 3×3 cm2 , 5×5 cm2 , 8×8 cm2 , 10×10 cm2 , and 20×20 cm2 . We evaluated the area under the PDD curve (AUC) for the buildup region for both 6X and 6XFFF beams. Ten SBRT lung and five IMRT sinuses plans were generated using the EclipseTM planning system, commissioned with AAA. Plan evaluation involved the comparison of the isodose distributions between plans of the two delivery modes. RESULTS: For small field sizes, the dose in the buildup region is greater for FFF beam than for flattened beam. The percentage differences between 6XFFF and 6X flattened beams for the following field sizes (2×2 cm2 , 3×3 cm2 , 5×5 cm2 , 8×8 cm2 , 10×10 cm2 , and 20×20 cm2 ) are 3.4%,2.2%, 7.2%, 4.9%, 0.9%, and -0.5%, respectively. Plan evaluation of the IMRT sinus cases found approximately 3% higher maximum dose within the PTV for FFF plans. No observable differences were found between two types of SBRT lung plans. CONCLUSIONS: For small fields, the dose buildup region of FFF beam is slightly greater than that of flattened beams. This effect is more pronounced in IMRT sinus cases, leading to higher dose within the PTV. There was no difference in the SBRT lung plans between the two beam modes.

Efecto del inhibidor de fosfodiesterasa tipo 4 —rolipram, sobre la maduración in vitro de oocitos bovinos / Efeito do inibidor de fosfodiesterasa tipo 4 —rolipram, sobre a maturação in vitro de oocitos bovinos / Effect of phosphodiesterase type 4-inhibitor rolipram on in vitro maturatrion of bovine oocytes

Durante el proceso de maduración in vitro de oocitos, las gonadotropinas FSH y LH inducen la producción de AMPc. El AMPc tiene efecto dual, donde los altos niveles de AMPc intraoocitario mantienen su bloqueo meiótico, mientras que en las células de la granulosa inducen la maduración del oocito. Los niveles de AMPc son regulados por hidrólisis mediada por fosfodiesterasas (PDE), las cuales presentan expresión específica en el folículo, el oocito expresa la PDE 3, mientras que las células de la granulosa PDE 4. Con el objetivo de evaluar el efecto del rolipram, un inhibidor específico de PDE 4, sobre el porcentaje de maduración nuclear in vitro (MNIV) de oocitos bovinos, 629 complejos cúmulo oocito (CCO) fueron cultivados a 38.5 ºC/5% CO2/24 h, en medio TCM-199 con la adición de pFSH y hrLH, o rolipram. Los grupos experimentales fueron: adición de gonadotropinas, rolipram (25, 50 ó 75 M), rolipram 50 M + gonadotropinas, o control sin estímulo. Los oocitos fueron teñidos con DAPI y evaluados bajo fluorescencia para determinar el porcentaje de maduración nuclear por la expulsión del primer cuerpo polar. El rolipram 50M estimuló la maduración nuclear de oocitos bovinos de una manera similar a la obtenida con las gonadotropinas (76.83 vs 79.46%, p>0.05), pero en mayor medida que la observada con rolipram 25 y 75 M (31.25, y 28.61%, respectivamente). Los CCOs cultivados en presencia de rolipram 50 M+Gonadotropinas maduraron en menor proporción (63.74%) comparada con gonadotropinas (p<0.01) o rolipram 50 M (p<0.05). Los resultados permiten concluir que el porcentaje de maduración nuclear in vitro de oocitos bovinos depende de la dosis de rolipram utilizada, donde la concentración de rolipram 50 M presentó un comportamiento similar a las gonadotropinas en la maduración del oocito...

Gonadotropic follicle stimulating hormone (FSH) and lutenizing hormone (LH) induce intracellular cyclic AMP (cAMP) production during the in vitro maturation (IVM) of bovine oocytes. Cyclic AMP exerts a dual effect, where high intraoocyte cAMP levels are responsible for oocyte meiotic blockage, while high cAMP levels into the granulose cells induce oocyte maturation. Intracellular cAMP levels are regulated by phosphodiesterases (PDE)-mediated hydrolysis, enzymes having a specific follicle expression pattern. Oocyte expresses typo 3 PDE (PDE 3), while granulose cells expresses type 4 PDE (PDE 4). With the aim to test the effect of the specific PDE 4 inhibitor rolliprom on percentage in vitro nuclear maturation (IVNM) of bovine oocytes, 629 cumulus-oocyte complexes (COC) were cultured at 38.5 ºC/CO2 5%/24 h on TCM-199 medium with pFSH and hrLH with or without rolipram. Experimental groups were: gonadotrophins alone, gonadotropins + rolipram (25, 50, or 75 M), rolipram 50 M + gonadotrophins, and control (media without stimulus). In order to determinate the nuclear maturation percentage by the first polar body expulsion, oocytes were dyed with DAPI and evaluated by fluorescence. Rolipram 50 M stimulated bovine oocyte nuclear maturation in a similar way to gonadotrophins stimulus (76.83 vs. 79.46%, p>0.05) did, but in a higher way than rolipram 25 M (31.25%) or 75 M (28.61%). The COC cultured with rolipram 50 M+gonadotrophins maturated in a lower proportion (63.74%) than did with gonadotropins (p<0.01) or rolipram 50 M (p<0.05). A dose-dependent response of percentage of IVNM of bovine oocytes was detected. Thus rolipram 50 M, exerts a similar effect of gonadotropins on oocyte maturation...

Durante o processo de maturação in vitro de oocitos, as gonadotrofinas FSH e LH induzem a produção de AMPc. O AMPc tem duplo efeito, pois os altos níveis de AMPc intraoocitario mantém o bloqueio meiótico, enquanto que nas células da granulosa induzem a maturação do oocito. Os níveis de AMPc são regulados pela hidrólise mediada das fosfodiesterasas (PDE), as quais apresentam expressão específica no folículo, o oocito expressa a PDE 3, enquanto que as células da granulosa PDE 4. Com o objetivo de avaliar o efeito do rolipram, um inibidor específico de PDE 4, sobre a percentagem de maturação nuclear in vitro (MNIV) de oocitos bovinos, 629 complexos cúmulo oocito (CCO) foram cultivados a 38.5 ºC/5% CO2/24 h, em meio TCM-199 com a adição de pFSH e hrLH, o rolipram. Os grupos experimentais foram: adição de gonadotrofinas, rolipram (25, 50 ó 75 M), rolipram 50 M + gonadotrofinas, ou controle sem estímulo. Os oocitos foram tingidos com DAPI e avaliados sob fluorescência para determinar a percentagem de maturação nuclear pela expulsão do primeiro corpo polar. O rolipram 50M estimulou a maturação nuclear de oocitos bovinos de maneira similar a obtida com as gonadotrofinas (76.83 vs 79.46%, p>0.05), porém em maior medida que a observada com rolipram 25 y 75 M (31.25, y 28.61%, respectivamente). Os CCOs cultivados na presença de rolipram 50 M+Gonadotrofinas maturaram em menor proporção (63.74%) quando comparado com gonadotrofinas (p<0.01) ou rolipram 50 M (p<0.05). Os resultados permitem concluir que a percentagem de maturação nuclear in vitro de oocitos bovinos depende da doses de rolipram utilizada, sendo que a concentração de rolipram 50 M apresentou um comportamento similar às gonadotrofinas na maturação do oocito...

Patterns of breast cancer relapse.

AIMS: Although breast cancer is the major cause of cancer-related death in women, there is little comprehensive information on long-term outcomes, particularly pertaining to site of relapse. The Strathfield Breast Centre (TSBC) is a multidisciplinary breast clinic that has collected patient data prospectively over 14 years. METHODS: All women with invasive, non-metastatic breast cancer, referred to TSBC from 1989 until 2002, were studied (n=2509). After initial treatment, patients were reviewed at 3-12-month intervals, including annual mammography and/or breast ultrasound. Information was collected on demographics, pre- and post-operative management and patient outcomes. Survival was analysed by the method of Kaplan and Meier. RESULTS: The mean age was 58 years and median follow-up 4 years (range <1-14) with complete data for 81%. In total, 456 patients (18%) had a local, nodal or distant relapse. The most common site of first relapse was to bone (in 125 patients), followed by local recurrence (124), lung (73) and liver (57). The median interval from primary breast surgery until recurrence was 2.3 years and disease-free intervals correlated to survival (p<0.0001). After local recurrence the 5-year survival was 41%, vs. 20% for nodal and 13% for distant recurrence (p<0.0001). Following breast-conserving surgery, the 5-year disease-free survival after local recurrence was 49.4%, vs. 33.1% after chest wall recurrence (p=0.0361). Of distant relapses, bone metastases had the best prognosis, with median survival 2.4 years. CONCLUSION: These data provide information on treatment outcomes in a multidisciplinary setting and statistical information that will be useful when discussing the fears and expectations of patients after the diagnosis of breast cancer.

Enhanced tissue strength in cryopreserved, collagen-based blood vessel constructs.

Traditional cryopreservation methods do not adequately preserve complex natural or engineered multicellular tissues due to the ice formation in the extracellular matrices. Vitrification is an alternate ice-free method for cryopreservation. This study compares the effects of vitrification and conventional cryopreservation on an engineered blood vessel construct. Collagen-based vascular constructs were used as models in this study. Tubular constructs were cut into rings and distributed into fresh, frozen, and vitrified groups for evaluation of mechanical properties and cell viability. Passive mechanical tests revealed enhanced tissue strength after both freezing and vitrification. Cryosubstitution studies of frozen and vitrified constructs revealed negligible ice in the vitrified specimens and extensive ice formation in the extracellular matrix of frozen specimens. Morphological changes associated with ice formation were visible within tissues preserved using traditional cryopreservation but not in tissue preserved using vitrification. The metabolic assay results indicated that vitrified tissue had similar viability to fresh controls. These results suggest that the increased tissue strength after cryopreservation may relate to thermal property change during preservation that cross-link collagen in tissue-engineered blood vessels. Further development of this cryopreservation method is necessary to minimize the alteration in material property and maintain cell viability of the constructs.

KI-67 AND hTERT expression can aid in the distinction between malignant and benign pheochromocytoma and paraganglioma.

The clinical and histopathological distinction between benign and malignant pheochromocytomas and paragangliomas is difficult, and reliable diagnostic markers are lacking. Here we have evaluated the prognostic value of human telomerase reverse transcriptase (hTERT) gene expression detected by reverse transcription PCR (RT-PCR); telomerase activity (TA) measured by TRAP (telomeric repeat amplification protocol) assay; immunohistochemical staining for Ki-67/MIB-1; and the mRNA expression of matrix metalloproteinase (MMP)-2 and EMMPRIN (extracellular matrix metalloproteinase inducer) analyzed by in situ hybridization in 32 primary pheochromocytomas or abdominal paragangliomas. hTERT was expressed in 7/11 malignant tumors (defined as presence of metastasis and/or extensive local invasion) as compared with in 2/21 benign tumors. All of the benign tumors showed <1% proliferative activity, as measured by Ki-67/MIB-1 staining. In all three patients with malignant tumors who developed metastases and/or invasive local recurrence during follow-up, the tumors were positive for either hTERT expression or Ki-67/MIB-1 immunoreactivity. TA was not a significant discriminator between benign and malignant tumors, and the value of EMMPRIN and MMP-2 as predictive markers was limited. In conclusion, the findings imply that the combined use of Ki-67/MIB-1 and hTERT, in addition to histopathology, provides a highly specific tool to identify benign pheochromocytoma and abdominal paraganglioma cases that are not at risk of developing recurrent or metastatic disease.

The management of benign and malignant pheochromocytoma and abdominal paraganglioma.

AIMS: To report treatment and outcome in patients with malignant and benign pheochromocytoma and abdominal paraganglioma. METHODS: Review of clinical and therapeutic features in 85 patients with pheochromocytoma or abdominal paraganglioma between 1976 and 1999. RESULTS: Thirty-nine of 85 patients presented with symptoms other than classical paroxysmal attacks. Paragangliomas were more often malignant (7/15) than pheochromocytomas (7/70). No recurrences have occurred in 71 patients with tumours initially classified as benign after a median follow-up time of 144 months (range 7-287). Nine of 14 patients with tumours classified as malignant have developed metastasis and/or local recurrence. Treatment of malignant tumours with cyclophosphamide, vincristine and dacarbazine (CVD) improved or stabilised the disease in three of four patients. CONCLUSION: Life long follow-up of patients with benign pheochromocytoma is not necessary. Combination chemotherapy (CVD) is a valid option in the treatment of malignant pheochromocytomas and abdominal paragangliomas.

Serological analysis of patients treated with a new surgical hemostat containing bovine proteins and autologous plasma.

A randomized, controlled clinical study of the management of diffuse bleeding with CoStasis surgical hemostat, a new hemostat containing bovine thrombin and collagen with the patient's own plasma, included patients undergoing cardiac, hepatic, iliac, and general surgery. Sera from 92 patients treated with CoStasis and 84 control patients were collected preoperatively and at a post surgical follow-up of 8 weeks. Among the control group, 57 patients were treated with Instat collagen sponge in noncardiac indications. Results showed that antibody responses in the CoStasis clinical study were similar to the reported literature for all antigens screened and were not associated with any adverse reactions. The bovine thrombin preparations in CoStasis and other commercially available thrombins were compared with the use of SDS-PAGE and Western blot analyses. Within this clinical study, CoStasis was shown to be a safe and effective hemostatic product containing bovine thrombin and bovine collagen and no pooled human blood products.

Serial [18F] fluorodeoxyglucose positron emission tomography after human neuronal implantation for stroke.

OBJECTIVE: There is no known effective treatment for chronic stroke. In this report, we used positron emission tomography (PET) with [18F]fluorodeoxyglucose (FDG) to map the metabolic brain response to neuronal cell implantation in the first human neuroimplantation trial for stroke. METHODS: Twelve patients (nine men, three women; mean age +/- standard deviation, 60.8+/-8.3 yr) with chronic basal ganglia infarction and persistent motor deficit underwent FDG PET within 1 week before and 6 and 12 months after stereotactic implantation of human neuronal cells. Serial neurological evaluations during a 52-week postoperative period included the National Institutes of Health stroke scale and the European stroke scale. RESULTS: Alterations in glucose metabolic activity in the stroke and surrounding tissue at 6 and 12 months after implantation correlated positively with motor performance measures. CONCLUSION: FDG PET performed as part of an initial open-label human trial of implanted LBS-Neurons (Layton BioScience, Sunnyvale, CA) for chronic stroke demonstrates a relationship between relative regional metabolic changes and clinical performance measures. These preliminary findings suggest improved local cellular function or engraftment of implanted cells in some patients.