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PubMed2004

Lumpectomy margins are affected by tumor size and histologic subtype but not by biopsy technique.

OBJECTIVE: The effect of the type of biopsy (needle vs. excisional) on lumpectomy margin status has not been well established. The objective of this study was to determine whether needle biopsy is associated with a higher positive margin rate at time of lumpectomy. METHODS: We evaluated this hypothesis in the setting of a prospective multi-institutional study. A total of 3975 patients were enrolled in the University of Louisville Breast Cancer Sentinel Lymph Node Study from May 7, 1998 to June 3, 2003. Patients who underwent lumpectomy at the time of their sentinel lymph node biopsy were the focus of this analysis. Patients with clinical stage T1 N0 and T2 N0breast cancer were eligible; 29 patients were found to have T3 tumors on final pathology. Pathologists at each institution defined margin positivity, and tumor at the inked margin of resection was the study guideline. RESULTS: Median patient age was 59 years, and median tumor size was 1.5 cm. A total of 2658 patients underwent lumpectomy with the following results. The cancer of 1515 patients was diagnosed by fine-needle or core-needle biopsy and of 821 patients was diagnosed by excisional biopsy; in 322 patients the method of diagnosis was unknown. The type of previous biopsy did not significantly affect the positive-margin rate at the time of lumpectomy (13.3% vs. 11.0% for needle and excisional biopsy, respectively, P = 0.107). However, patients with larger tumors were more often found to have a positive margin (11.4% vs. 13.9% vs. 27.6% for T1, T2, and T3 tumors, respectively; P = 0.010). No difference was found in margin status after excision of palpable versus nonpalpable tumors (10.6% vs. 10.9%, respectively, P = 0.743). Histologic subtype, however, did affect margin status (15.8% vs. 9.8% positive margins for lobular vs. ductal type, respectively, P = 0.003). CONCLUSIONS: In this multi-institutional study, increasing tumor size and lobular histologic subtype were associated with a greater likelihood of a positive margin. The type of biopsy, needle or excisional, had no effect on the ability to achieve negative margins.

Adult↗
PubMed2005

Local control in ductal carcinoma in situ treated by excision alone: incremental benefit of larger margins.

BACKGROUND: Margin width has been shown previously to be the most important predictor of local treatment failure after breast conservation for ductal carcinoma in situ (DCIS). METHODS: Five variables thought to be associated with local recurrence were evaluated by univariate and multivariate analysis in 455 nonrandomized patients with DCIS treated with excision alone. RESULTS: Multivariate analysis showed that margin width, age, nuclear grade, and tumor size all were independent predictors of local recurrence, with margin width as the single most important predictor. After adjusting for all other predictors the likelihood of local recurrence for patients with margins less than 10 mm was 5.39 times as much as that for patients with margins of 10 mm or more (95% confidence interval, 2.68-10.64). CONCLUSIONS: Margin width, the distance between DCIS and the closest inked margin, was the single most important predictor of local recurrence. As margin width increases, the risk for local recurrence decreases.

Adult↗
PubMed2005

Understanding the mechanisms creating false positive lumpectomy margins.

OBJECTIVE: All breast surgeons deal with the frustration of initial pathologic close or positive margins that have no residual cancer upon re-excision. To understand the mechanisms that create false positive margins, specimen handling was standardized in a single surgeon's practice and margin issues were tracked. METHODS: Prospectively over a 3.5-year period, needle-localized lumpectomies for the management of early-stage breast cancer were standardized in all aspects of specimen handling, including surgeon inking and specimen compression for specimen radiography for quality assurance. The current study reviews 220 such cases where the original lumpectomy included a small piece of overlying skin from over the target lesion. All specimen radiography was performed with compression at the skin to deep level to bias the "pancaking" effect of pushing tumor to specimen surface to the deep margin. RESULTS: Of the 220 therapeutic lumpectomies performed for clinical stage 0-2 breast cancer in this fashion, 175 (79.5%) had negative margins by a distance of 10 mm or more. Margins less than 10 mm were classified as close and were present in 20 (9.1%) of cases. These were heavily biased toward margins closer than 2 mm. Positive initial margins accounted for 25 (11.4%) of cases. Of the 45 close or positive margin cases, 12 involved the deep margin only, and on re-excision none was found to have residual tumor. When other single margins were involved, re-excision found tumor in 5 of 14 cases (35.7%). When multiple margins were close or positive, 9 of 19 cases (47.3%) were found to have residual tumor at re-excision. CONCLUSIONS: Specimen compression increases the incidence of false margin positivity. The best predictors of true margin positivity are multiple close or positive margins or margin positivity in a direction not associated with specimen ex vivo compression.

Breast Neoplasms↗
PubMed2006

The influence of additional surgical margins on the total specimen volume excised and the reoperative rate after breast-conserving surgery.

BACKGROUND: It is unclear whether the additional removal of breast tissue during breast-conserving therapy (BCT) for breast cancer beyond the standard lumpectomy reduces the incidence of inadequate microscopic margins found at pathological examination and subsequent reoperation. This study compares the reoperative rates after initial BCT in 3 groups of patients who underwent lumpectomy with complete resection of 4 to 6 additional margins, lumpectomy with selective resection of 1 to 3 additional margins, or standard lumpectomy. METHODS: Retrospective data were reviewed from 171 selected cases of BCT, from May 2000 to February 2006. Forty-five cases involved lumpectomy with complete resection of 4 to 6 additional margins; 77 involved lumpectomy with selective resection of 1 to 3 additional margins, whereas 49 involved standard lumpectomy. All samples underwent pathologic analysis of inked resection margins by permanent section. The 3 groups were compared for patient demographics, tumor size and histologic subtype, tumor stage, margin status, excised specimen volume, and eventual subsequent reoperation. Adequate surgical margin was defined as any negative margin greater than 2 mm. RESULTS: The group with complete resection of 4 to 6 additional margins had a subsequent reoperation rate of 17.7%, whereas the group with selective resection of 1 to 3 additional margins and the standard lumpectomy group had a subsequent reoperation rate of 32.5% and 38.7%, respectively, because of inadequate margins. The mean total excised specimen volume in the 3 groups was 129.19, 46.04, and 37.44 cm3, respectively. CONCLUSIONS: The complete resection of 4 to 6 additional margins during the initial BCT resulted in the lowest subsequent reoperation rate, and the largest total volume specimen excised among the 3 techniques studied.

Adult↗
PubMed2005

Effects of unilateral naris closure on the nasal and maxillary sinus mucosa in rabbit.

In nasal septal deviation, the deviated narrow side is more frequently combined with sinusitis due to the mechanical obstruction of ostiomeatal unit. The purpose of this study was to determine whether reduced airflow has a harmful effect on nasal inflammation and the development of rhinosinusitis. Twenty-three healthy New Zealand white rabbit were used. One side of the nostril was closed in 15 rabbits and 5 rabbits served as control. Rabbits were sacrificed at 4, 8 and 12 weeks. The anterior maxillary and septal mucosa were obtained for EM and LM study. Mucociliary clearance rate was measured using the India ink method. At 4 weeks, the opened side mucosa showed ciliary losses and ulcerations in the mucosa, but at 8 and 12 weeks, the closed side showed more severe ulceration and ciliary loss than open side. The mucociliary clearance rate was 12.3 mm/min in the control group, but the open side was 8.7 and 7.1 mm/min and the closed side was 4.7 and 6.4 mm/min at 4 and 8 weeks, respectively. Electron microscopy showed many abnormal cilia at 8 weeks in both sides. These observations suggest that inadequate airflow is an important causative factor of nasal and sinus infection, and that patients with septal deviation have a similar chance of developing sinusitis in either side.

Animals↗
PubMed2006

[Bacterial meningitis in HIV patient: think about co-infections!].

The authors report the case of an African 34-year-old patient who was admitted to the intensive care unit for bacterial meningitis due to Streptococcus pneumoniae. A meningeal co-infection due to Cryptococcal neoformans was found the 3rd day in an HIV infection context. Cryptococcus neoformans detection in cerebrospinal fluid, using the India-ink stain, has a low sensibility which imposes the search of cryptococcal antigen and the culture of cerebrospinal fluid. These last two exams have a sensibility of at least 90%.

Adult↗
PubMed2006

Preferred viewing distance and screen angle of electronic paper displays.

This study explored the viewing distance and screen angle for electronic paper (E-Paper) displays under various light sources, ambient illuminations, and character sizes. Data analysis showed that the mean viewing distance and screen angle were 495 mm and 123.7 degrees. The mean viewing distances for Kolin Chlorestic Liquid Crystal display was 500 mm, significantly longer than Sony electronic ink display, 491 mm. Screen angle for Kolin was 127.4 degrees, significantly greater than that of Sony, 120.0 degrees. Various light sources revealed no significant effect on viewing distances; nevertheless, they showed significant effect on screen angles. The screen angle for sunlight lamp (D65) was similar to that of fluorescent lamp (TL84), but greater than that of tungsten lamp (F). Ambient illumination and E-paper type had significant effects on viewing distance and screen angle. The higher the ambient illumination was, the longer the viewing distance and the lesser the screen angle. Character size had significant effect on viewing distances: the larger the character size, the longer the viewing distance. The results of this study indicated that the viewing distance for E-Paper was similar to that of visual display terminal (VDT) at around 500 mm, but greater than normal paper at about 360 mm. The mean screen angle was around 123.7 degrees, which in terms of viewing angle is 29.5 degrees below horizontal eye level. This result is similar to the general suggested viewing angle between 20 degrees and 50 degrees below the horizontal line of sight.

Adolescent↗
PubMedDate not supplied

The new fabrication method of standard surface sources.

We developed a new fabrication method for standard surface sources by using an inkjet printer with inks in which a radioactive material is mixed to print on a sheet of paper. Three printed test patterns have been prepared: (1) 100mm x 100mm uniformity-test patterns, (2) positional-resolution test patterns with different widths and intervals of straight lines, and (3) logarithmic intensity test patterns with different radioactive intensities. The results revealed that the fabricated standard surface sources had high uniformity, high positional resolution, arbitrary shapes and a broad intensity range.

Calibration↗
PubMed2004

Split-line pattern and histologic analysis of a human osteochondral plug graft.

The split-line pattern of collagen fibers in articular cartilage is oriented according to weight-bearing stresses. The importance of aligning the split-line pattern of articular cartilage in human osteochondral plug grafts relative to the surrounding cartilage has been proposed but not evaluated clinically. The purpose of this case report is to compare the articular split-line pattern with the histologic analysis of a specimen obtained from a patient who underwent an osteochondral plug transfer procedure with subsequent conversion to joint arthroplasty. The fresh osteochondral section obtained at total knee arthroplasty, including the area treated 18 months earlier with an osteochondral plug transfer, was fixed in formalin. Split-lines were demonstrated on the surface using a needle dipped in India ink. After decalcification and paraffin embedding, the specimen was sectioned and stained with Safranin O and fast green for histologic analysis. The split-line pattern of the anterior portion of the graft was oriented parallel to the resident cartilage; however, collagen orientation was divergent (approximately 30 degrees oblique) to the surrounding cartilage near the posterior portion of the graft. It was at this margin that further resident articular degeneration was noted despite the plug articular surface remaining relatively intact. Although bony incorporation of the plug occurred, there was a residual chondral cleft at the graft-host interface. Our report suggests that an osteochondral plug transfer can survive despite a slightly divergent collagen split-line pattern of plug relative to the resident articular bed. Perhaps more important to the clinical outcome is the selection of true focal, traumatic lesions rather than lesions degenerative in etiology, no matter how focal they appear.

Aged↗
PubMed2005

Vascularity of the hip labrum: a cadaveric investigation.

PURPOSE: The purpose of this study was to define the vascularity of the hip labrum and to identify regional differences in vascular penetration that may have implications for the healing potential of this structure. TYPE OF STUDY: Injection study of human cadavers to investigate the vascularity of the hip labrum. METHODS: Twelve hips from 6 human frozen cadavers devoid of severe articular pathology were used. High-resolution surface-coil magnetic resonance imaging (MRI) was performed in both sagittal and coronal planes to better define anatomic planes in 2 dimensions and to correlate Spalteholz sections with the surrounding joint and labral anatomy. Each pelvis was injected with intra-arterial India Ink and frozen; 3-mm sagittal or coronal sections were then cut and processed using a modified Spalteholz technique, yielding anatomic zones of the labrum. Six specimens were cut in the sagittal plane and 6 were cut in the coronal plane. Specimens were examined at x10 magnification with transillumination. Anatomic zones with regional variations in vascularity were defined. RESULTS: The anterior and superior aspects of the labrum showed degenerative changes on MRI and under direct manual transillumination in 75% of specimens. Overall, there was a relatively poor vascular supply to the labrum; however, there were regional differences between anatomic zones. Zone I (capsular contribution) had significantly more vascularity than zone II (articular side) (P < .01). Zone IA (the portion of the zone not attached to bone) showed the most consistent source of vessels across all specimens (smallest variation between specimens); however, zone IB (the portion of the zone attached to bony acetabulum) had the greatest overall mean vascularity score. These differences were not statistically significant. Furthermore, vascularity patterns were not significantly different in the anterior, superior, posterior, and inferior labral regions, nor were they different in torn versus intact specimens. CONCLUSIONS: The cadaveric specimens evaluated in this study had a relatively avascular hip labrum. However, the increased vascularity seen in zone I (capsular side) may have implications for treatment, similar to that described in the meniscus of the knee. CLINICAL RELEVANCE: A better understanding of the vascularity of the hip labrum will guide treatment of labral pathology and may have implications for the healing potential of this structure. Labral tears occurring in the vascular zone may be amenable to arthroscopic repair rather than debridement.

Acetabulum↗
PubMed2005

Split-line orientation of the talar dome articular cartilage.

PURPOSE: The purpose of this study was to define the pattern of superficial collagen fibril orientation on the talar dome and to create a template for rotational graft alignment during mosaicplasty. TYPE OF STUDY: An anatomic, cadaveric study. METHODS: Eight fresh-frozen human cadaveric tali had the articular surface of the talar dome pierced in grid fashion with a dissecting needle dipped in India ink. The longitudinal split that occurred between the superficial collagen fibers defined the fiber arrangement at each site of the talar dome. RESULTS: There was a remarkably consistent pattern to the split lines from specimen to specimen. CONCLUSION: Proper orientation of mosaicplasty plugs during transfer from the donor to recipient site with respect to superficial collagen fiber arrangement may allow the graft to better resist tensile stresses and may contribute to increased long-term graft longevity. CLINICAL RELEVANCE: Use of the talar dome map, combined with knowledge of the split-line orientation of the donor site, should allow surgeons performing mosaicplasty procedures on the talus to properly rotate their plugs with respect to split-line orientation.

Aged↗
PubMed2003

Virus particle detection by solid phase immunocapture and atomic force microscopy.

A novel application of atomic force microscopy (AFM) in the rapid, label-free detection and identification of viruses is described. Multiplexed, miniaturized antibody domains were constructed using "ink-jet" protein arraying technology. The solid-phase affinity substrate termed the "ViriChip" was used in the immunocapture of bacteriophage fd, canine parvoviruses, and coxsackieviruses and analyzed by AFM. Immunocapture was found to be antibody-specific with a sensitivity of 10(8)pfu/ml in 30min. Virus binding was found to be linear for concentration between 10(8) and 10(10)pfu/ml and did not reach saturation through 4h.

Equipment Design↗
PubMed2004

Roles of a conserved family of adaptor proteins, Lnk, SH2-B, and APS, for mast cell development, growth, and functions: APS-deficiency causes augmented degranulation and reduced actin assembly.

Lnk, SH2-B, and APS form a conserved adaptor protein family. All of those proteins are expressed in mast cells and their possible functions in signaling through c-Kit or FcRI have been speculated. To investigate roles of Lnk, SH2-B or APS in mast cells, we established IL-3-dependent mast cells from Ink-/-, SH2-B-/-, and APS -/- mice. IL-3-dependent growth of those cells was comparable. Proliferation or adhesion mediated by c-Kit as well as degranulation induced by cross-linking FcRI were normal in the absence of Lnk or SH2-B. In contrast, APS-deficient mast cells showed augmented degranulation after cross-linking FcRI compared to wild-type cells, while c-Kit-mediated proliferation and adhesion were kept unaffected. APS-deficient mast cells showed reduced actin assembly at steady state, although their various intracellular responses induced by cross-linking FcRI were indistinguishable compared to wild-type cells. Our results suggest potential roles of APS in controlling actin cytoskeleton and magnitude of degranulation in mast cells.

Actins↗
PubMed2006

An investigation of genomic base distribution.

While veritable oceans of ink have been spilled over the base distributions within genes, the literature is virtually silent on large scale intra genomic base distribution. To address this issue, we have examined approximately 3400 chromosomal sequences from approximately 2000 entire genomes-including DNA and RNA, single- and double-stranded, coding and non-coding genomes. For each sequence the mean, variance, skewness, and kurtosis for each base were computed along with the genome base composition. The main findings are: (1) there is no simple relationship between these statistics and the base composition of the genome, (2) in non-viral genomes, base distribution is non-uniform, (3) base distribution in non-eukaryotic genomes obeys a number of simple rules, (4) these rules are not dependent on the presence of coding sequences, (5) bacterial genomes in particular are unusually compliant with these rules, and (6) eukaryotes have a unique pattern of base distribution.

Base Composition↗
PubMed2006

Electrochemical detection of E. coli 16S rDNA sequence using air-plasma-activated fullerene-impregnated screen printed electrodes.

A new DNA modified electrode for the electrochemical detection of 16S rDNA extracted from Escherichia coli (JCM1649) is proposed. The electrodes were fabricated by screen printing a fullerene-impregnated carbon ink onto a poly(methylmethacrylate) substrate and immobilizing a probe DNA on the surface after activating the electrode with air plasma. The results indicated a dramatic improvement in the surface coverage of the immobilized probe DNA, and of the reduction peak of the redox indicator (Co(phen)(3)(3+)) due to the incorporation of fullerene. By immobilizing the probe onto the fullerene-impregnated screen-printed electrodes, the PCR product of the 16S rDNA extracted from E. coli was directly detected without any pretreatment. A well defined signal difference was observed between the perfectly matching oligonucleotide and the mismatching one, and it was possible to detect the target at the modified electrode. This method enabled us to clearly detect the two base mismatches in the ca. 1500-bases long 16S rDNA sequence.

Air↗
PubMed2005

Micropatterned polymer substrates control alignment of proliferating Schwann cells to direct neuronal regeneration.

Microcontact printed polymeric substrates were evaluated for their ability to control Schwann cell attachment and direct proliferation, as Schwann cell guidance is a crucial factor in directing peripheral nerve regeneration. Elastomeric stamps of poly(dimethylsiloxane) were "inked" with laminin, a permissive protein for Schwann cell adhesion, and stamped onto poly(methyl methacrylate) substrates to create patterns of lines and intervals varying from 10 to 50 microm wide. Schwann cells were seeded onto the substrates in serum-free media. After 4h, media was replaced with serum-containing growth media and changed daily thereafter. The addition of growth media to stimulate proliferation initially caused some loss in cell orientation relative to the laminin pattern, but when monolayer formation was complete, a high degree of cell orientation was observed. As both cell-cell contacts and surface coverage were maximized, the Schwann cells achieved an even higher order of orientation than observed during the early stages of proliferation. Significantly, smaller pattern widths increased the degree of orientation, regardless of interval width. Our results indicate that patterned polymeric substrates may enhance peripheral nerve regeneration by creating a highly ordered Schwann cell matrix for guidance of neurons.

Adsorption↗
PubMed2005

Direct patterning of mammalian cells onto porous tissue engineering substrates using agarose stamps.

This paper describes simple, inexpensive, and potentially generic methodology for generating patterns of mammalian cells on porous scaffolds for tissue engineering using replica printing. Circular patterns (diameter: 200, 700, and 1000 microm) of human osteoblasts were transferred directly from topographically patterned agarose stamps onto porous hydroxyapatite scaffolds or onto fibronectin-coated glass slides. The use of hydrogel stamps provided a "wet", biocompatible surface and maintained the viability of cells adsorbed on stamps during the patterning process. Stamps inked once with suspensions of cells allowed the repeated patterning of substrates. Direct stamping of human osteoblasts (and, potentially other mammalian cells) can be used to control the size, spacing, and geometry of patterns of cells printed on porous tissue engineering substrates. This approach may find use in controlling the spatial invasion of scaffolds, promoting the hierarchical organization of cells, and in controlling cell-cell interactions as a step in preservation of phenotypes of cells.

Actins↗
PubMed2005

Stem cell-coated titanium implants for the partial joint resurfacing of the knee.

The goal of the present study was to evaluate the partial surface replacement of the knee with stem cell-coated titanium implants and to provide a basis for a successful treatment of large osteochondral defects. Mesenchymal stem cells (MSCs) were isolated from bone marrow aspirates of adult sheep. Round titanium implants with a diameter of 2 x 7.3 mm were seeded with autologous MSC and inserted into an osteochondral defect in the medial femoral condyle. As controls, defects received either an uncoated implant or were left untreated. Nine animals with 18 defects were sacrificed after 6 months. Histological evaluation was performed by intravital polychrome fluorescent labelling, intravital perfusion with Indian ink, microradiographs and differential staining with toluidine blue. The quality of regenerated cartilage was assessed by in situ hybridization of collagen type II and immunohistochemistry of collagen types I and II. In 50% of the cases, defects treated with MSC-coated implants showed a complete regeneration of the subchondral bone layer. In these cases collagen type II and only traces of collagen type I were detected. A high level of collagen type II mRNA expression compared to articular cartilage indicates regenerating hyaline-like cartilage. A total of 50% of MSC-coated and uncoated implants failed to osseointegrate and formation of fibrocartilage was observed. Untreated defects as well as defects treated with uncoated implants demonstrated incomplete healing of subchondral bone and formation of fibrous cartilage. A modified histological score according to Wakitani significantly demonstrated better results for cell-coated implants (8.8+/-6.4) than for uncoated implants (5.5+/-3.9) and for untreated defects (2.8+/-2.5). Our results demonstrate that, in a significant number of cases, a partial joint resurfacing of the knee with stem cell-coated titanium implants occur. A slow bone and cartilage regeneration and an incomplete healing in half of the MSC-coated implants are limitations of the presented method. To improve our approach and optimize the experimental parameters, further investigations are needed prior to clinical application.

Animals↗