Extent of surgical procedure in triple negative breast carcinomas.

INTRODUCTION: Triple negative breast carcinomas (TNBC) account for approximately 15-20% of all breast carcinomas. This subtype is characterised by an unfavourable prognosis with early locoregional recurrence a metastases. Only few studies have focused on the impact of local surgery on the overall therapeutic outcome. However, decisions are difficult to make in the case of TNBC, and no particular molecular subtype or marker exists that would make the decision-making process easier. The aim of our retrospective study was to analyse the TNBC surgical management outcomes at EUC Clinic in Zlin. METHODS: 440 women with breast carcinoma were operated on at EUC Clinic from 2014 to 2016, including 29 patients with TNBC; bilateral carcinoma was present in one case. Neoadjuvant chemotherapy (NAC) was indicated in 6 cases. The tumour centre was marked with a clip. The extent of surgery depended on the residual size of the tumour. Sentinel lymph node biopsy was indicated in clinically negative lymph nodes; further management followed the Z0011 study if the biopsy was positive. Axillary lymph node dissection was performed after NAC. In all cases, surgery was followed by systemic chemotherapy, and by radiotherapy in the case of breast-conserving procedures. RESULTS: The group included 29 women and one patient with bilateral carcinoma, i.e. 30 cases of TNBC. Mean age was 57 years and median age was 55.5 years. Mean follow-up was 62.9 months, with the median of 69.9 month. NAC was indicated in 6 patients; complete pathological response was achieved in one case. NAC was followed by mastectomy in 5 cases including a bilateral procedure in one case, and by breast-conserving surgery in one case. Axillary dissection was performed in all cases. Breast-conserving surgery and sentinel node biopsy predominated in the group (16 cases). Local recurrence was observed in 4 cases, 2 times as an isolated local recurrence after one year and 2 times as part of generalization, always after mastectomy. Six patients died of generalized disease. No regional recurrence was observed. CONCLUSION: TNBC is characterised by a worse prognosis and a higher rate of local recurrence. As confirmed by our study, the results of breast-conserving surgery can be comparable to those of radical procedures, and thus radical surgery should be indicated prudently.

Ductal carcinoma in situ and the extent of surgery.

INTRODUCTION: Ductal carcinoma in situ (DCIS) is a very heterogenous disease. The incidence of DCIS has been increasing with the adoption of mammography screening. This opened new questions concerning surgical and adjuvant therapy.  Methods: We retrospectively observed the incidence of DCIS amongst the patients that underwent surgical resection in EUC clinic Zlín between 2017 and 2019. We also assessed the extent of breast surgery including interventions in axilla and the adjuvant therapy.  Results: There were 616 breast cancer patients, of whom 44 (7.1%) were diagnosed with DCIS. Breast-conserving surgery was performed in 35 (80%) patients. Lumpectomy alone was performed in 21 (47%) patients. Mastectomy was indicated primarily in 9 cases with additional two mastectomies performed to achieve clear margins. All sentinel nodes were negative.  Conclusion: Results confirmed, that the surgical therapy as well as radiotherapy and hormonal treatment are performed according to guidelines at our department. Proportion of sentinel node biopsy is remarkably higher, therefore an improvement in this area is our next goal.

Targeted axillary dissection with preoperative tattooing of biopsied positive axillary lymph nodes in breast cancer.

Sentinel lymph node biopsy (SLNB) has emerged as an alternative to axillary lymph node dissection during breast cancer surgery during the last 2 decades. However, there are several controversies regarding the indication of the sentinel node biopsy after neoadjuvant chemotherapy which can convert positive lymph nodes to negative. The false-negative rate after neoadjuvant chemotherapy is unacceptably high. This high false-negative rate can be decreased by marking of the positive lymph nodes and removal during sentinel lymph node biopsy procedure in addition to the sentinel lymph nodes. The aim of this study was to investigate the possibility of carbon tattooing of the positive sentinel lymph nodes before neoadjuvant chemotherapy. In 2016, a prospective protocol was launched investigating the black carbon tattooing procedure of the suspective and positive axillary lymph nodes by injecting 0.1-0.5 carbon ink in normal saline under ultrasound guidance. All patients underwent black carbon tattooing of the suspected or positive axillary lymph nodes before the chemotherapy or one week before the primary surgery when chemotherapy was not indicated in the neoadjuvant setting. Sentinel lymph nodes together with lymph nodes marked by the black carbon ink were removed and histologically evaluated. So far 27 patients were treated under this protocol. Breast saving surgery was performed in 22 cases and mastectomy in 5 cases. All patients had invasive ductal carcinoma. In 20 patients neoadjuvant chemotherapy was indicated and in 7 patients primary surgery was performed. All lymph nodes marked by black carbon ink were successfully identified and removed. Sentinel lymph node biopsy was performed in 8 cases and sentinel lymph node biopsy followed by axillary dissection in 15 cases. Axillary dissection alone was performed in 4 cases. In 19 cases, the black carbon ink was present in the sentinel lymph node at the same time and in 4 cases carbon dye was present in other lymph nodes than the lymph node identified during SLNB, which corresponds to 17.4%. In the group of patients undergoing primary surgery, in one case from six, the sentinel lymph node was negative and the lymph node marked with carbon ink positive which represents false-negative lymph node and failure of the SLNB procedure. After neoadjuvant chemotherapy, there was no false-negative lymph node identified, but the conversion of the positive lymph nodes to negative was present in 10 cases (50%). There were no complications attributed to carbon ink tattooing. The results of positive sentinel lymph nodes tattooing have confirmed that this method is safe and allows a decrease in the false negativity rate during the sentinel node biopsy procedure.

In-focus electron microscopy of frozen-hydrated biological samples with a Boersch phase plate.

We report the implementation of an electrostatic Einzel lens (Boersch) phase plate in a prototype transmission electron microscope dedicated to aberration-corrected cryo-EM. The combination of phase plate, C(s) corrector and Diffraction Magnification Unit (DMU) as a new electron-optical element ensures minimal information loss due to obstruction by the phase plate and enables in-focus phase contrast imaging of large macromolecular assemblies. As no defocussing is necessary and the spherical aberration is corrected, maximal, non-oscillating phase contrast transfer can be achieved up to the information limit of the instrument. A microchip produced by a scalable micro-fabrication process has 10 phase plates, which are positioned in a conjugate, magnified diffraction plane generated by the DMU. Phase plates remained fully functional for weeks or months. The large distance between phase plate and the cryo sample permits the use of an effective anti-contaminator, resulting in ice contamination rates of <0.6 nm/h at the specimen. Maximal in-focus phase contrast was obtained by applying voltages between 80 and 700 mV to the phase plate electrode. The phase plate allows for in-focus imaging of biological objects with a signal-to-noise of 5-10 at a resolution of 2-3 nm, as demonstrated for frozen-hydrated virus particles and purple membrane at liquid-nitrogen temperature.

Sequence, genomic structure, and chromosomal assignment of human DOC-2.

DOC-2 is a human gene originally identified as a 767-bp cDNA fragment isolated from normal ovarian epithelial cells by differential display against ovarian carcinoma cells. We have now determined the complete cDNA sequence of the 3.2-kb DOC-2 transcript and localized the gene to chromosome 5. A 12.5-kb genomic fragment at the 5'-end of DOC-2 has also been sequenced, revealing the intron-exon structure of the first eight exons (788 bases) of the DOC-2 gene. Translation of the DOC-2 cDNA predicts a hydrophobic protein of 770 amino acid residues with a molecular weight of 82.5 kDa. Comparison of the DNA and amino acid sequences of DOC-2 to publicly accessible sequence databases revealed 83% identify to p96, a murine protein of similar size, thought to be a mitogen-responsive phosphoprotein. In addition, about 45% identity was observed between the first 140 N-terminal residues of DOC-2 and the Caenorhabditas elegans M110.5 and Drosophila melanogaster Dab genes.

Isolation of a gene (DLG3) encoding a second member of the discs-large family on chromosome 17q12-q21.

The discs-large family is a collection of proteins that have a common structural organization and are thought to be involved in signal transduction and mediating protein-protein interactions at the cytoplasmic surface of the cell membrane. The defining member of this group of proteins is the gene product of the Drosophila lethal (1) discs large (dlg) 1 locus, which was originally identified by the analysis of recessive lethal mutants. Germline mutations in dlg result in loss of apical-basolateral polarity, disruption of normal cell-cell adhesion, and neoplastic overgrowth of the imaginal disc epithelium. We have isolated and characterized a novel human gene, DLG3, that encodes a new member of the discs-large family of proteins. The putative DLG3 gene product has a molecular weight of 66 kDa and contains a discs-large homologous region, a src oncogene homology motif 3, and a domain with homology to guanylate kinase. The DLG3 gene is located on chromosome 17, in the same segment, 17q12-q21, as the related gene, DLG2. The products of the DLG2 and DLG3 genes show 36% identity and 58% similarity to each other, and both show nearly 60% sequence similarity to p55, an erythroid phosphoprotein that is a component of the red cell membrane. We suggest that p55, DLG2, and DLG3 are closely related members of a gene family, whose protein products have a common structural organization and probably a similar function.

Isolation and mapping of a gene encoding a novel human ADP-ribosylation factor on chromosome 17q12-q21.

A gene encoding a low-molecular-weight GTP-binding protein was isolated from a retinal cDNA library and mapped to human chromosome 17q12-q21. Comparison of the predicted protein with the protein databases revealed striking homology to the family of ADP-ribosylation factors (ARFs), which are thought to be involved in membrane trafficking and protein secretion. The greatest homology observed was with the rat ARF-like 4 protein (ARL4), with which it shared 58% identity, while the more highly conserved human ARF1 and ARF3 proteins each shared 46% identity. Inspection of the predicted new protein showed that it contained each of the six conserved motifs that are required for guanine nucleotide binding and hydrolysis, and thus it is probably a novel ARF isoform. We have designated the new protein and its corresponding gene ARF4L.

A strategy for constructing high-resolution genetic maps of the human genome: a genetic map of chromosome 17p, ordered with meiotic breakpoint-mapping panels.

Genetic linkage analyses with genotypic data obtained from four CEPH reference families initially assigned 24 new PCR-based markers to chromosome 17 and located the markers at specific intervals of an existing genetic map of chromosome 17p. Each marker was additionally genotyped with an ordered set of obligate, phase-known recombinant chromosomes. The breakpoint-mapping panels for each family consisted of two parents, one sib with a nonrecombinant chromosome, and one or more sibs with obligate recombinant chromosomes. The relative order of markers was determined by sorting segregation patterns of new markers and ordered anchor markers and by minimizing double-recombination events. Consistency of segregation patterns with multiple flanking loci constituted support for order. A genetic map of chromosome 17p was completed with 39 markers in 23 clusters, with an average space of 3 cM between clusters. The collection of informative genotypes was highly efficient, requiring fivefold fewer genotypes than would be collected with all the CEPH families. Given the availability of large numbers of highly informative PCR-based markers, meiotic breakpoint mapping should facilitate construction of a human genomic map with 1-cM resolution.

A PCR-based genetic map for human chromosome 3.

Oligonucleotide primers for 125 simple sequence repeat microsatellite-based genetic markers have been assayed by polymerase chain reaction (PCR) in the CEPH reference family panel. These microsatellites include 101 dinucleotide repeats as well as 24 new tetranucleotide repeats. The average heterozygosity of this marker set was 72.4%. Genetic data were analyzed with the genetic mapping package LINKAGE. A subset of these microsatellite markers define a set of 56 uniquely ordered loci (> 1000:1 against local inversion) that span 271 cM. Sixty-seven additional loci were tightly linked to markers on the uniquely ordered map, but could not be ordered with such high precision. These markers were positioned by CMAP into confidence intervals. One hundred thirteen of the microsatellite markers were also tested on a chromosome 3 framework somatic cell hybrid panel that divides this chromosome into 23 cytogenetically defined regions, integrating the genetic and physical maps of this chromosome. The high density, high heterozygosity, and PCR format of this genetically and physically mapped set of markers will accelerate the mapping and positional cloning of new chromosome 3 genes.

[Late recurrence of female genital tuberculosis]. / Spätrezidive der weiblichen Genitaltuberkulose

5 years patient-material: 336 cases (1042 case-sheats) were analysed for the purpose of studying the cases with relapse. 28 recurrence--7,62 per cent--were registered. Relapse is regarded, if the properly treated patient remained well for at least 5 years, without any signs and symptoms and after negative check-ups carried out at regular intervals became actively ill again. If active disease ocurred earlier this was not regarded a relapse but rather the progression of the original disease, which could be explained either by drug-resistance or insufficient conservative treatment.