Inflammatory breast cancer defined: proposed common diagnostic criteria to guide treatment and research.

PURPOSE: Inflammatory breast cancer is a deadly and aggressive type of breast cancer. A key challenge relates to the need for a more detailed, formal, objective definition of IBC, the lack of which compromises clinical care, hampers the conduct of clinical trials, and hinders the search for IBC-specific biomarkers and treatments because of the heterogeneity of patients considered to have IBC. METHODS: Susan G. Komen, the Inflammatory Breast Cancer Research Foundation, and the Milburn Foundation convened patient advocates, clinicians, and researchers to review the state of IBC and to propose initiatives to advance the field. After literature review of the defining clinical, pathologic, and imaging characteristics of IBC, the experts developed a novel quantitative scoring system for diagnosis. RESULTS: The experts identified through consensus several "defining characteristics" of IBC, including factors related to timing of onset and specific symptoms. These reflect common pathophysiologic changes, sometimes detectable on biopsy in the form of dermal lymphovascular tumor emboli and often reflected in imaging findings. Based on the importance and extent of these characteristics, the experts developed a scoring scale that yields a continuous score from 0 to 48 and proposed cut-points for categorization that can be tested in subsequent validation studies. CONCLUSION: To move beyond subjective 'clinical diagnosis' of IBC, we propose a quantitative scoring system to define IBC, based on clinical, pathologic, and imaging features. This system is intended to predict outcome and biology, guide treatment decisions and inclusion in clinical trials, and increase diagnostic accuracy to aid basic research; future validation studies are necessary to evaluate its performance.

The staining potential of various currently marketed mouthrinses.

OBJECTIVES: The objectives of this clinical trial were to determine the tooth staining potential as measured by the Macpherson Modification of the Lobene Stain Index, and degree of taste alteration of four currently marketed mouthrinses when used over a 12-week period. METHODS: This investigation consisted of a 12-week, observer-blind, single-center, randomized comparison of five parallel groups of subjects. One-hundred and seventy-one subjects granting their informed consent completed the trial. Subjects were randomized to one of four currently marketed mouthrinses Crest PRO-HEALTH Rinse (CPH), Cepacol (C), Scope (S), Viadent ADVANCED CARE (V), or brushing alone (BA) with a currently marketed fluoride toothpaste. Upon randomization, subjects received a baseline stain score and then a prophylaxis to remove all extrinsic stain. Clinical assessments were repeated after six weeks and three months of product use, and subjects were asked to complete a questionnaire after the first use, at day 4, day 14, at six weeks, and 12 weeks to assess potential taste alteration. RESULTS: CPH and C demonstrated significantly (p < 0.001) more extrinsic stain after six weeks of use, and CPH, C (p < 0.001), and S (p = 0.01) after 12 weeks of use versus brushing alone with fluoride toothpaste. V was not significantly different from brushing alone at either time point. After six weeks of using the product as directed, up to 53% of subjects using CPH experienced taste interference for up to three hours post-rinse. CONCLUSIONS: The results of this study demonstrated that regular use of CPH and C mouthrinses resulted in extrinsic stain accumulation after six weeks, with increased accumulation after 12 weeks versus brushing alone.

Vaginal progesterone for prevention of preterm birth in asymptomatic high-risk women with a normal cervical length: a systematic review and meta-analysis.

Objective: To determine whether vaginal progesterone reduces spontaneous preterm birth (sPTB) before 37 weeks in asymptomatic high-risk women with a singleton pregnancy and normal mid-gestation cervical length.Study design: Databases were searched (from inception to December 2020) with the search terms "progesterone" and "premature birth" or "preterm birth". Studies were screened and included if they assessed vaginal progesterone compared to placebo in women with normal cervical length. Data were pooled and synthesized in a meta-analysis using a random effects model.Data sources: MEDLINE and Embase databases.Study synthesis: Following PRISMA screening guidelines, data from 1127 women across three studies were available for synthesis. All studies had low risk of bias and were of high quality. The primary outcome was sPTB <37 weeks, with secondary outcomes of sPTB <34 weeks. Vaginal progesterone did not significantly reduce sPTB before 37 weeks, or before 34 weeks with a relative risk (RR) of 0.76 (95% CI 0.37-1.55, p = .45) and 0.51 (95% CI 0.12-2.13, p = .35), respectively.Conclusions: Vaginal progesterone does not decrease the risk of sPTB in high-risk singleton pregnancies with a normal mid-gestation cervical length.

Targeting GLI1 expression in human inflammatory breast cancer cells enhances apoptosis and attenuates migration.

BACKGROUND: Inflammatory breast cancer (IBC) is an aggressive subtype of breast cancer with distinct molecular profiles. Gene expression profiling previously identified sonic hedgehog (SHH) as part of a gene signature that is differentially regulated in IBC patients. METHODS: The effects of reducing GLI1 levels on protein expression, cell proliferation, apoptosis and migration were determined by immunoblots, MTT assay, Annexin-V/PI assay and conventional and automated cell migration assays. RESULTS: Evaluation of a panel of breast cancer cell lines revealed elevated GLI1 expression, typically a marker for hedgehog-pathway activation, in a triple-negative, highly invasive IBC cell line, SUM149 and its isogenic-derived counterpart rSUM149 that has acquired resistance to ErbB1/2 targeting strategies. Downregulation of GLI1 expression in SUM149 and rSUM149 by small interfering RNA or a small molecule GLI1 inhibitor resulted in decreased proliferation and increased apoptosis. Further, GLI1 suppression in these cell lines significantly inhibited cell migration as assessed by a wound-healing assay compared with MCF-7, a non-invasive cell line with low GLI1 expression. A novel high-content migration assay allowed us to quantify multiple effects of GLI1 silencing including significant decreases in cell distance travelled and linearity of movement. CONCLUSION: Our data reveal a role for GLI1 in IBC cell proliferation, survival and migration, which supports the feasibility of targeting GLI1 as a novel therapeutic strategy for IBC patients.

Host-interactive genes in Amerindian Helicobacter pylori diverge from their Old World homologs and mediate inflammatory responses.

Helicobacter pylori is the dominant member of the gastric microbiota and has been associated with an increased risk of gastric cancer and peptic ulcers in adults. H. pylori populations have migrated and diverged with human populations, and health effects vary. Here, we describe the whole genome of the cag-positive strain V225d, cultured from a Venezuelan Piaroa Amerindian subject. To gain insight into the evolution and host adaptation of this bacterium, we undertook comparative H. pylori genomic analyses. A robust multiprotein phylogenetic tree reflects the major human migration out of Africa, across Europe, through Asia, and into the New World, placing Amerindian H. pylori as a particularly close sister group to East Asian H. pylori. In contrast, phylogenetic analysis of the host-interactive genes vacA and cagA shows substantial divergence of Amerindian from Old World forms and indicates new genotypes (e.g., VacA m3) involving these loci. Despite deletions in CagA EPIYA and CRPIA domains, V225d stimulates interleukin-8 secretion and the hummingbird phenotype in AGS cells. However, following a 33-week passage in the mouse stomach, these phenotypes were lost in isolate V225-RE, which had a 15-kb deletion in the cag pathogenicity island that truncated CagA and eliminated some of the type IV secretion system genes. Thus, the unusual V225d cag architecture was fully functional via conserved elements, but the natural deletion of 13 cag pathogenicity island genes and the truncation of CagA impaired the ability to induce inflammation.

An RNA polymerase-binding protein that is required for communication between an enhancer and a promoter.

Although bacteriophage T4 late promoters are selectively recognized by Escherichia coli RNA polymerase bearing a single protein encoded by T4 gene 55 (gp55), efficient transcription at these promoters requires enhancement by the three T4 DNA polymerase accessory proteins, bound to distal "mobile enhancer" sites. Two principles are shown to govern this transcriptional enhancement: (i) Promoter recognition and communication between the enhancer and the promoter require separate phage-coded proteins. Only RNA polymerase that has the T4 gene 33 protein (gp33) bound to it is subject to enhancement by the three DNA replication proteins. (ii) Transcriptional enhancement in this prokaryotic system is promoter-specific. Promoter specificity is generated by a direct competition of phage T4 gp33 and gp55 with the E. coli promoter recognition protein, sigma 70, for binding to the E. coli RNA polymerase core. Thus, polymerase that contains sigma 70 is competent to transcribe T4 early and middle genes, but lacks the ability to be enhanced by the DNA replication proteins, while polymerase that contains gp55 and gp33 is capable of enhancement via gp33, but its activity is restricted to T4 late promoters by gp55.

Sizing criteria for a low footprint passive mine water treatment system.

The objective of this paper is to present data from a novel vertical flow mine water treatment system, demonstrate how these data can be used to generate sizing formulae for this technology, and present a comparison between the size of system based on these formulae and those of conventionally designed passive systems. The paper focuses on passive treatment of circum-neutral ferruginous mine waters bearing up to 50 mgl(-1) of iron in either ferrous or ferric form. The Vertical Flow Reactor (VFR) operates by passing mine water down through an accreting bed of ochre, the ochre bed being responsible for the intensification of iron removal by self-filtration and/or autocatalytic iron oxidation and precipitation. Key to the design and operation of the VFR system is the decrease in permeability in this ochre bed over time. The paper demonstrates that the VFR system can remove iron at many times the 10 g/m2/day removal rate - an often employed figure for the sizing of aerobic settling ponds and wetlands. The paper demonstrates that VFRs are viable and novel passive treatment system for mine waters with a smaller footprint than conventional systems.

Affinity-based screening of combinatorial libraries using automated, serial-column chromatography.

We have developed an automated serial chromatographic technique for screening a library of compounds based upon their relative affinity for a target molecule. A "target" column containing the immobilized target molecule is set in tandem with a reversed-phase column. A combinatorial peptide library is injected onto the target column. The target-bound peptides are eluted from the first column and transferred automatically to the reversed-phase column. The target-specific peptide peaks from the reversed-phase column are identified and sequenced. Using a monoclonal antibody (3E-7) against beta-endorphin as a target, we selected a single peptide with sequence YGGFL from approximately 5800 peptides present in a combinatorial library. We demonstrated the applicability of the technology towards selection of peptides with predetermined affinity for bacterial lipopolysaccharide (LPS, endotoxin). We expect that this technology will have broad applications for high throughout screening of chemical libraries or natural product extracts.

Comparative biological responses to human Sonic, Indian, and Desert hedgehog.

A comprehensive comparison of Sonic (Shh), Indian (Ihh), and Desert (Dhh) hedgehog biological activities has not previously been undertaken. To test whether the three higher vertebrate Hh proteins have distinct biological properties, we compared recombinant forms of the N-terminal domains of human Shh, Ihh, and Dhh in a variety of cell-based and tissue explant assays in which their activities could be assessed at a range of concentrations. While we observed that the proteins were similar in their affinities for the Hh-binding proteins; Patched (Ptc) and Hedgehog-interacting protein (Hip), and were equipotent in their ability to induce Islet-1 in chick neural plate explant; there were dramatic differences in their potencies in several other assays. Most dramatic were the Hh-dependent responses of C3H10T1/2 cells, where relative potencies ranged from 80nM for Shh, to 500nM for Ihh, to >5microM for Dhh. Similar trends in potency were seen in the ability of the three Hh proteins to induce differentiation of chondrocytes in embryonic mouse limbs, and to induce the expression of nodal in the lateral plate mesoderm of early chick embryos. However, in a chick embryo digit duplication assay used to measure polarizing activity, Ihh was the least active, and Dhh was almost as potent as Shh. These findings suggest that a mechanism for fine-tuning the biological actions of Shh, Ihh, and Dhh, exists beyond the simple temporal and spatial control of their expression domains within the developing and adult organism.

Regular articles: conditional disruption of hedgehog signaling pathway defines its critical role in hair development and regeneration.

Members of the vertebrate hedgehog family (Sonic, Indian, and Desert) have been shown to be essential for the development of various organ systems, including neural, somite, limb, skeletal, and for male gonad morphogenesis. Sonic hedgehog and its cognate receptor Patched are expressed in the epithelial and/or mesenchymal cell components of the hair follicle. Recent studies have demonstrated an essential role for this pathway in hair development in the skin of Sonic hedgehog null embryos. We have further explored the role of the hedgehog pathway using anti-hedgehog blocking monoclonal antibodies to treat pregnant mice at different stages of gestation and have generated viable offspring that lack body coat hair. Histologic analysis revealed the presence of ectodermal placode and primodium of dermal papilla in these mice, yet the subsequent hair shaft formation was inhibited. In contrast, the vibrissae (whisker) development appears to be unaffected upon anti-hedgehog blocking monoclonal antibody treatment. Strikingly, inhibition of body coat hair morphogenesis also was observed in mice treated postnatally with anti-hedgehog monoclonal antibody during the growing (anagen) phase of the hair cycle. The hairless phenotype was reversible upon suspension of monoclonal antibody treatment. Taken together, our results underscore a direct role of the Sonic hedgehog signaling pathway in embryonic hair follicle development as well as in subsequent hair cycles in young and adult mice. Our system of generating an inducible and reversible hairless phenotype by anti-hedgehog monoclonal antibody treatment will be valuable for studying the regulation and mechanism of hair regeneration.

Two universally conserved adenosines of the group I intron that are important for self-splicing but not for core catalytic activity.

Two adenosine residues, universally conserved among group I introns, are located in the L4 region of the catalytic core. Base-substitution mutations in these adenosines resulted in diminished in vitro self-splicing activity of the Tetrahymena group I intron, more severely for double than for single mutations. The defect caused by the mutation of the L4 adenosines was manifest at the first step of splicing (cleavage of the 5' splice site by a guanosine molecule), and could be overcome by increasing the magnesium ion concentration of the reaction buffer. In contrast, a related activity of the group I intron, specific hydrolysis of the 3' splice site, was virtually unaffected by the L4 adenosine mutations; this reaction must require an active conformation of the catalytic site of the ribozyme and also depends on the recognition of guanosine (the guanosine residue that precedes the 3' splice site in all group I introns). These results suggest that the role of the L4 adenosines is limited to improving the reactivity of the 5' splice site during splicing. We also found that mutations that eliminate the 5' splice site-bearing P1 stem and the P10 stem have little or no effect on specific hydrolysis of the 3' splice site.

Peripartum changes in maternal cerebral blood flow velocity in normotensive and preeclamptic patients.

OBJECTIVE: To compare the effect of delivery on cerebral blood flow velocity between normotensive and preeclamptic women, adjusting for mode of delivery and change in hematocrit values. METHODS: Twenty-three normotensive and 46 preeclamptic women had maternal middle cerebral blood flow velocity assessed antepartum and at 24 and 48 hours postpartum. In addition, hematocrit changes and method of delivery were recorded. We then compared the effect of delivery on cerebral blood flow velocity changes using analysis of variance with Student t test for significance. RESULTS: Cerebral blood flow velocity was significantly higher in preeclamptic than in normotensive women (P < .05) and rose significantly in the postpartum period (P < .004). Neither hematocrit change nor mode of delivery affected cerebral blood flow velocity changes. CONCLUSION: Cerebral blood flow velocity showed minimal peripartum changes in the normotensive group but increased significantly postpartum in preeclamptic women in a setting of minimal arterial pressure change.

Changes in cerebral perfusion pressure in puerperal women with preeclampsia.

OBJECTIVE: To determine the variation in the estimated maternal cerebral perfusion and cerebrovascular resistance (the resistance area product) in the puerperium. METHODS: The maternal middle cerebral artery was evaluated by transcranial Doppler ultrasound in ten women 2 days before labor, in 21 women in early labor and at 24 and 48 hours postpartum, and in 6 women at 1 week postpartum. Cerebral blood flow velocities were determined. Women were diagnosed initially with mild preeclampsia. Estimated cerebral perfusion pressure was Vmean/[Vmean - Vdiastolic] [BPmean - BPdiastolic]. Because the diameter of the vessels could not be measured directly, an index of resistance was calculated: the resistance area product = BPmean/velocitymean. We calculated an index of cerebral blood flow to be estimated cerebral perfusion pressure divided by resistance area product. Our study had a power of 80% to detect a 16-cm/second increase in middle cerebral blood flow velocity. RESULTS: Estimated maternal cerebral perfusion was maintained for up to 1 week postpartum. Cerebrovascular resistance did not change in the puerperium. Cerebral blood flow index (+/-standard deviation) was significantly increased at 1 week postpartum compared with early labor levels (28.3 +/-6.9 versus 46.7+/-15.6, respectively) (P < .05). CONCLUSION: Cerebral blood flow 1 week postpartum increased significantly over early labor values. These persistent changes in the cerebral vasculature might put patients at risk for seizures up to 1 week postpartum.

The late luteal phase in infertile women: comparison of simultaneous endometrial biopsy and progesterone levels.

Endometrial biopsy specimens were obtained from 107 normally menstruating infertile women 2 to 3 days before the anticipated onset of menses and were day-dated according to histologic criteria. A simultaneous blood sample was obtained for measurement of progesterone (P) and beta-subunit of human chorionic gonadotropin. Of 98 biopsies which could be accurately dated, 56 were in-phase (IP) and 42 were out-of-phase (OOP). Mean serum P levels were significantly lower in women with OOP biopsies undertaken more than 4 days before the onset of menses. A sharp decline in serum P levels was observed in women with IP but not OOP biopsies, so that on the final premenstrual day serum P levels were significantly higher than normal in women with OOP biopsies. Pregnancy continued without interruption in two of six patients who underwent biopsy in the cycle of conception. One patient had an ectopic pregnancy; and the three remaining pregnant patients, all with subnormal P values, aborted. The study suggests that there is a high frequency of minor abnormalities in luteal function in normally menstruating, infertile women for whom tubal and male factors were normal. The frequency of subclinical pregnancy (2 of 107) was lower than anticipated from earlier studies.

Antepartum middle mean cerebral blood flow velocity correlation with maternal hemodynamics.

OBJECTIVE: To determine the correlation between simultaneous assessment of maternal middle cerebral blood flow velocity with the other maternal hemodynamic factors of cardiac output and mean arterial pressure. STUDY DESIGN: Eight normotensive patients were assessed. Maternal cerebral blood flow velocity was assessed using transcranial Doppler. Cardiac output was assessed noninvasively using the thoracic electrical bioimpedance technique over four cycles. Transcranial assessment of cerebral blood flow velocity was done over four cycles. Statistical analysis was then done using the Pearson correlation coefficient and linear regression analysis with stepwise regression. A p-value of < 0.05 was considered significant. RESULTS: The value of the hemodynamic parameters were cardiac output 8.6 +/- 2.6 L/min, mean arterial pressure 82 +/- 9.7 mm Hg, and mean maternal cerebral blood flow velocity 59.6 +/- 11 cm/s. The pulsatility index was 0.85 +/- 0.15. The mean blood pressure could only explain 42% of the variation in systolic maternal cerebral blood flow velocity and 32% of the variation in mean maternal cerebral blood flow velocity. The mean middle cerebral blood flow velocity did not correlate with cardiac output. CONCLUSIONS: Middle cerebral artery velocity correlates moderately with mean arterial pressure but not with cardiac output. The control of mean arterial pressure cannot be used as the only indicator of appropriate reduction in cerebral blood flow velocity.

A multi-element analysis of the effect of teacher aides in an "open"-style classroom.

Open classrooms with few rules, individualized instruction, and informal class organization present new problems for the application of behavior principles. The effects of three types of teacher aides on student achievement and on-task behavior were studied. Each was compared with a standard no-aide condition. Subjects were 54 third graders in two "open"-style classrooms. The three types of aide, helping adult, disciplinary adult, and helping fifth-grade aide, were compared in a multi-element design with a no-aide control. The helping-adult aide significantly affected the academic output of the class, when compared with the no-aide condition. All aide conditions produced more academic work and on-task behavior than did the standard no-aide condition.

A retrospective analysis of invasive Streptococcus pneumoniae infections in Trinidad.

A retrospective analysis of culture-positive cases of S pneumoniae from normally sterile body fluids is reported. Over 40% of patients were 5 years old or less while 28% of patients were 50 years old or more. Meningitis (44%) was the commonest clinical presentation followed closely by pneumonia (31%). The commonest predisposing disorder was human immunodeficiency virus infection though there were no identifiable risk factors in the majority of patients. Mortality from invasive pneumococcal disease was significantly higher in elderly patients compared with other age groups (p = 0.0003). In this study, all S pneumoniae isolates, for which there were antibiotic sensitivity data, were penicillin and/or amoxycillin sensitive.

Human stress protein hsp70: overexpression in E coli, purification and characterization.

The gene encoding the stress-inducible member of human heat shock protein hsp70, was expressed in E. coli using the bacteriophage T7 RNA polymerase-based gene expression system. Recombinant hsp70 (R-hsp70) was purified from inclusion bodies after solubilization and refolding, using a combination of ATP-agarose affinity chromatography and ion-exchange chromatography. R-hsp70 was shown to be monomeric and free of its structurally similar E. coli counterpart, DnaK. In addition, R-hsp70 is functional as demonstrated by its ability to bind to peptides and to ATP. The availability of pure, correctly folded R-hsp70 in sufficient quantity will assist in the structural and functional characterization of hsp70. Furthermore, an understanding of the cytoprotective function of hsp70 and its role in immune responses during infections will be facilitated by the availability of pure R-hsp70.