Aspirin/antiplatelet agent use improves disease-free survival and reduces the risk of distant metastases in Stage II and III triple-negative breast cancer patients.

PURPOSE: The objective is to define the therapeutic role of antiplatelet agents in a triple-negative breast cancer (TNBC) population. METHODS: We performed retrospective analysis using the UTSW TNBC registry containing data from 222 Stage II-III TNBC patients treated between 1998 and 2016. Univariate analysis and multivariable logistic regression models were constructed to identify factors associated with disease-free survival (DFS), distant metastases rate (DMR), and overall survival outcomes. Antiplatelet drug use was determined by review of electronic medical records. RESULTS: A total of 65 patients used antiplatelet (AP) agents, and 157 patients did not use AP agents. Median follow-up for AP and non-AP groups was 41.3 and 40.9 months, respectively. There was an improvement in the AP group compared with the control group in 5-year DFS (80.4% at 5 years compared with 62.3% in the control group, p = 0.04) and 5-year DMR (8.8 vs. 31.9%, p = 0.007). In multivariate analysis, AP use was found to be significantly associated with improvements in DFS and DMR. CONCLUSIONS: We illustrate that antiplatelet agent use improves DMR and DFS among a stage II and III TNBC population despite our short follow-up evaluation. Longer follow-up evaluation will be required to determine additional outcome advantage for antiplatelet agent use. Our findings support consideration of investigation of antiplatelet therapy as an adjunctive therapy for TNBC at high risk for disease recurrence.

A systematic analysis of cell cycle regulators in yeast reveals that most factors act independently of cell size to control initiation of division.

Upstream events that trigger initiation of cell division, at a point called START in yeast, determine the overall rates of cell proliferation. The identity and complete sequence of those events remain unknown. Previous studies relied mainly on cell size changes to identify systematically genes required for the timely completion of START. Here, we evaluated panels of non-essential single gene deletion strains for altered DNA content by flow cytometry. This analysis revealed that most gene deletions that altered cell cycle progression did not change cell size. Our results highlight a strong requirement for ribosomal biogenesis and protein synthesis for initiation of cell division. We also identified numerous factors that have not been previously implicated in cell cycle control mechanisms. We found that CBS, which catalyzes the synthesis of cystathionine from serine and homocysteine, advances START in two ways: by promoting cell growth, which requires CBS's catalytic activity, and by a separate function, which does not require CBS's catalytic activity. CBS defects cause disease in humans, and in animals CBS has vital, non-catalytic, unknown roles. Hence, our results may be relevant for human biology. Taken together, these findings significantly expand the range of factors required for the timely initiation of cell division. The systematic identification of non-essential regulators of cell division we describe will be a valuable resource for analysis of cell cycle progression in yeast and other organisms.

A flexible quantitative methodology for the analysis of gene-flow between conventionally bred maize populations using microsatellite markers.

Previous studies of gene-flow in agriculture have used a range of physical and biochemical markers, including transgenes. However, physical and biochemical markers are not available for all commercial varieties, and transgenes are difficult to use when trying to estimate gene flow in the field where the use of transgenes is often restricted. Here, we demonstrate the use of simple sequence repeat microsatellite markers (SSRs) to study gene flow in maize. Developing the first quantitative analysis of pooled SSR samples resulted in a high sampling efficiency which minimised the use of resources and greatly enhanced the possibility of hybrid detection. We were able to quantitatively distinguish hybrids in pools of ten samples from non-hybrid parental lines in all of the 24 pair-wise combinations of commercial varieties tested. The technique was used to determine gene flow in field studies, from which a simple model describing gene flow in maize was developed.

Epidermal growth factor induces the differential release of GP2 and amylase from AR4-2J cells.

Epidermal growth factor (EGF) stimulates secretion of glycoprotein 2 (GP2) in a time-and concentration-dependent manner from the AR4-2J pancreatoma cell line. Cell differentiation induced by dexamethasone treatment for 3 d, however, did not significantly alter either basal or EGF-stimulated GP2 release. Basal and EGF-stimulated GP2 release were similarly unaffected by caerulein, which promotes amylase secretion by a regulated route. A brief exposure to cycloheximide profoundly blocked EGF-evoked GP2 secretion. Furthermore, EGF-stimulated GP2 release was not accompanied by significant alterations in intracellular ionic calcium levels, in contrast to the stimulatory actions of caerulein. We conclude that EGF-stimulated release of GP2 occurs via a novel secretory pathway that is neither regulated nor constitutive as currently defined.

Use of alternative promoters to express the aromatase cytochrome P450 (CYP19) gene in breast adipose tissues of cancer-free and breast cancer patients.

Estrogen biosynthesis in adipose tissue has assumed great significance in terms of a number of estrogen-related diseases. Recent evidence suggests that estrogen synthesized locally in the breast is of singular significance in the development of breast cancer in elderly women. The biosynthesis of estrogen from C19 steroids is catalyzed by a specific form of cytochrome P450, namely aromatase cytochrome P450 (P450arom; the product of the CYP19 gene). The human CYP19 gene comprises nine coding exons, II-X, and its transcripts are expressed in the ovary, placenta, testes, adipose tissue, and brain. Tissue-specific expression of the CYP19 gene is determined, at least in part, by the use of tissue-specific promoters, which give rise to transcripts with unique 5'-noncoding termini. Transcripts in adipose tissue contain 5'-termini derived from specific untranslated exons, corresponding to expression derived from the proximal promoter II and its splice variant I.3, as well as a distal promoter, I.4. The object of the present study was to determine the distribution of these various exon-specific transcripts in breast adipose tissues from cancer-free women undergoing reduction mammoplasty and from patients with breast cancer, because this would provide important clues as to the nature of the factors regulating aromatase expression in these sites. To achieve this, we employed competitive RT-PCR, utilizing an internal standard for each exon-specific transcript of the CYP19 gene, as well as for the coding region, to evaluate total CYP19 gene transcripts. In cancer patients (n = 18), total CYP19 gene transcript levels were significantly higher in adipose tissue proximal to a tumor in comparison with adipose tissue distal to a tumor, in agreement with previous findings. Moreover, total transcript levels were higher in breast adipose tissue of cancer patients in comparison with those of cancer-free individuals (n = 9), even when the adipose tissue from the cancer patient was taken from a quadrant with no detectable tumor. We observed that exon I.4-specific transcripts were predominant in breast adipose obtained from cancer-free women. In this tissue, promoter-II-specific and exon I.3-specific transcripts were present in low copy number. On the other hand, in breast cancer patients, CYP19 gene transcripts from breast adipose tissue had primarily promoter-II-specific and exon I.3-specific sequence, whereas comparatively few transcripts had exon I.4-specific sequence at the 5'-terminus. We conclude that CYP19 gene transcription in breast adipose tissue of cancer-free individuals uses preferably promoter I.4, implicating a role of glucocorticoids and members of the IL-6 cytokine family in the regulation of this expression. On the other hand, the increased expression in breast adipose tissue bearing a carcinoma results from expression from promoters II and I.3, which are regulated by unknown factors acting via increased cAMP formation, which are presumably secreted by the tumor or associated cells.

Altered glucocorticoid rhythm attenuates the ability of a chronic SSRI to elevate forebrain 5-HT: implications for the treatment of depression.

Both glucocorticoids and selective serotonin reuptake inhibitors (SSRIs) alter aspects of 5-HT function including somatodendritic 5-HT1A autoreceptor sensitivity. Many depressed patients prescribed SSRIs have pre-existing flattened diurnal gluococorticoid rhythm. In these patients, interactions between flattened glucocorticoid rhythm and chronic SSRIs, which impact on the SSRI's ability to elevate forebrain 5-HT, may alter clinical efficacy. To address this issue rats underwent implantation of slow-release corticosterone (75 mg pellet s.c.) (to flatten the glucocorticoid rhythm) or sham surgery, and injection of fluoxetine (10 mg/kg/day i.p., 12 days) or vehicle. Using microdialysis in the frontal cortex we found that (21 h after the last injection) extracellular 5-HT was elevated in fluoxetine- or corticosterone-treated animals, but not in those treated with corticosterone plus fluoxetine. In fluoxetine-treated animals, blockade of terminal reuptake by local perfusion of fluoxetine increased 5-HT to the same level as it did in controls, suggesting normal terminal 5-HT release after chronic fluoxetine. However, 5-HT levels following local reuptake blockade in both the corticosterone and corticosterone plus fluoxetine groups were lower than controls, suggesting a corticosterone-induced decrease in terminal release. Finally in fluoxetine, corticosterone, and corticosterone plus fluoxetine groups, there was marked 5-HT1A receptor desensitization, evidenced by attenuation of the decrease in 5-HT release following systemic fluoxetine injection. The data indicate that, despite desensitization of 5-HT1A autoreceptors, concurrent flattened glucocorticoid rhythm compromises the ability of SSRIs to elevate forebrain 5-HT. These findings suggest a potential mechanism for the reduced antidepressant efficacy of SSRIs in those patients with pre-existing glucocorticoid abnormalities.

Acute and chronic effects of corticosterone on 5-HT1A receptor-mediated autoinhibition in the rat dorsal raphe nucleus.

Corticosteroid modulation of 5-HT(1A) receptor function may contribute to the aetiology of affective disorders. To examine this modulation, the effects of acute and chronic corticosterone administration on 5-HT(1A) autoreceptor function were investigated using in vitro electrophysiology in the rat dorsal raphe nucleus (DRN). The magnitude and time course of the inhibitory response to a submaximal dose of 5-HT was not affected by acute application of either corticosterone (30-200 nM) or dexamethasone (100 nM) in vitro, when tested either in slices from control rats or rats adrenalectomised two weeks prior to recording. For chronic treatment, rats were supplied with drinking water containing corticosterone (50 microg/ml) or ethanol vehicle (0.5%) for 25-31 days. The autoinhibitory response to 5-HT was significantly attenuated in the corticosterone-treated group; vehicle EC(50)=48+/-8 microM vs. corticosterone EC(50)=121+/-20 microM. Furthermore a subpopulation of 5-HT neurones from corticosterone-treated animals exhibited marked insensitivity to 5-HT. In situ hybridisation histochemistry showed that corticosterone did not affect the expression of mRNA encoding the 5-HT(1A) receptor or either the type 1 and type 3 subunits of the G-protein linked inwardly rectifying K+ (GIRK) channel. However, GIRK2 subunit mRNA expression was significantly reduced. Thus, 5-HT(1A) autoreceptor function in the DRN is attenuated following chronic, but not acute, exposure to elevated corticosterone levels, and this effect may involve changes to the receptor-effector coupling mechanism.

Serotonergic neurotransmission in the spinal cord and motor cortex of patients with motor neuron disease and controls: quantitative autoradiography for 5-HT1a and 5-HT2 receptors.

Serotonin 5-HT is a potent modulator of motor neuron excitability in the spinal cord. Serotonergic neurotransmission, because of its effects on glutamatergic excitation, may be relevant to the pathogenesis and therapy of motor neuron disease (MND). The human motor system was studied at two levels, spinal cord and motor cortex, by autoradiography for the 5-HT1A and 5-HT2 receptor subclasses. In addition, biochemical estimations of indole metabolites were performed in the spinal cord. Post mortem tissue from control cases and MND patients showed a reduction in 5-HT1A receptor binding in the cervical (p < 0.01) but not lumbar ventral horn in MND. 5-HT2 receptors were preserved in the ventral horn at both levels and were focally abundant around motor neuron somata. Tissue levels of 5-HT were unchanged in the spinal cord in MND. The metabolite 5-HIAA was increased in the cervical spinal cord in MND as was the molar ratio of 5HIAA:5-HT, implying that there may be an increased turnover of 5HT. In the motor cortex and premotor cortex the 5-HT1A receptor remained unchanged in MND. There was a 20% reduction in 5-HT2 receptor binding sites (p < 0.05) across all the cortical laminae with preservation of the normal pattern of laminar binding. These changes in two levels of the motor system in MND most likely represent physiological adaptations in the spinal cord and motor cortex rather than primary involvement of the serotonergic system in the pathogenesis of the disease.

Sentinel lymph node biopsy, an alternative to elective axillary dissection for breast cancer.

BACKGROUND: Axillary metastases remain an important prognostic indicator in breast cancer. Axillary lymphadenectomy (ALND) carries significant morbidity and is unnecessary in most patients with early breast cancer; thus, sentinel lymph node (SLN) biopsy has been advocated for axillary staging. We studied the SLN identification rate and its accuracy in predicting axillary metastases. METHODS: One hundred nineteen women with breast carcinoma underwent SLN and ALND. Lymphoscintigraphy was performed using Technetium99 sulfur colloid supplemented by Isosulfan blue dye. Hematoxylin/eosin-stained lymph node sections were examined by light microscopy. RESULTS: The SLN identification rate was 81%. One SLN was negative (1%) in a patient with axillary disease. SLN histology correctly predicted the absence of axillary disease in 98.6%. Sensitivity, specificity, and positive and negative predictive values were 96%, 100%, 100%, and 99%, respectively. CONCLUSIONS: Sentinel lymph node biopsy accurately predicts total axillary status and is valuable in the surgical staging of breast cancer.

Fine Needle Aspiration Cytology and Histologic Findings of Granular Cell Tumor of the Breast: Review of 19 Cases with Clinical/Radiologic Correlation.

Granular cell tumors (GCTs) are rare lesions in the breast of putative schwannian origin. These tumors are found in multiple sites throughout the body and have a characteristic histologic appearance. Recognition of these usually benign tumors is important since clinically, radiologically, and grossly GCTs of the breast often mimic carcinoma. The literature on these lesions in the breast is confined to isolated case reports. We describe the epidemiologic, cytologic, pathologic, and radiologic findings in 19 GCTs of the breast in 16 patients diagnosed at the University of Texas Southwestern Medical Center between 1991 and 1997.

Acute renal failure in six horses resulting from haemodynamic causes.

Six horses had been admitted to the hospital because of illness other than renal failure; diarrhoea, myositis, abdominal pain and/or suspected bacterial sepsis. Hypotension and disseminated intravascular coagulopathy were frequent findings in the horses. Abnormally high serum creatinine concentration and urine specific gravity of less than 1.022 were found in the horses with acute renal failure. Hyponatraemia and hypochloraemia were the most common abnormal electrolyte findings. Pronounced hyperkalaemia was not found. Variable degrees of tubular necrosis were seen in three of the four horses that had kidney sections submitted for microscopic examination. Renal cortical necrosis occurred in one horse. Intravenous fluid and electrolyte replacement was the most important therapy in those cases that were non-oliguric. Furosemide, mannitol and dopamine were used in horses with oliguria. The prognosis was generally good if the predisposing cause could be corrected and the acute renal failure was not oliguric.

Therapeutic use of gentamicin in horses: concentrations in serum, urine, and synovial fluid and evaluation of renal function.

Serum, synovial fluid, and urine concentrations of gentamicin were measured in normal mature horses which had been given a single dose of the drug. Mean peak serum concentration (16.8 microgram/ml) occurred in horses 30 minutes after they were given a single intramuscular dose of 4.4 mg of gentamicin/kg of body weight. In horses given a smaller dose of gentamicin (1.7 mg/kg), mean peak serum concentrations of gentamicin (10.2 microgram/ml) appeared at 1 hour. Synovial fluid concentration was maximum at 2 hours for both doses; in horses given the larger dose, mean peak concentration was 6.4 microgram/ml, and in those given the smaller dose (1.7 mg/kg), 3.4 microgram/ml. Measurable concentrations of gentamicin in serum and synovial fluid persisted 8 hours. During the first 8 hours, percentages of gentamicin excreted in the urine were between 3.9 and 32.8% of the larger dose and between 3.3 and 13.4% of the smaller dose. Serum creatinine concentrations were serially measured in 10 hospitalized horses intramuscularly given 1.7 to 4.4 mg of gentamicin/kg 4 times a day' significant increase in creatinine concentration was not found.

The AAMFT-Head Start Training Partnership Project: enhancing MFT capacities beyond the family system.

This paper summarizes the experience of a two-year project to promote partnerships between 12 graduate Marriage and Family Therapy (MFT) training programs and their 12 local Head Start partners. The American Association for Marriage and Family Therapy (AAMFT) awarded 9 grants to MFT-Head Start partnerships that had established relationships with each other prior to the project. Three MFT-Head Start pairs that had no preexisting relationship but wanted to develop one received planning grants. The partnership project provided an exciting opportunity for MFT faculty and interns to integrate family systems theory into larger organizational settings and to enhance the capacity of MFTs to serve disadvantaged, low-income, culturally diverse families and the community agencies that work with them. Preparing MFT students to intervene and respond systemically both in organizations and with the families they serve represents a viable alternative to more traditional approaches to MFT that limit the focus to the family system. Survey results and interview data reflect the rich experiences of MFT faculty and supervisors, MFT interns, and Head Start staff. Implementation issues are discussed and recommendations are made to improve the collaboration between MFTs and community-based agencies such as Head Start.

Malignant hyperthermia in a halothane-anesthetized horse.

Malignant hyperthermia developed in a 4-year-old Thoroughbred horse following 3 hours and 15 minutes of halothane anesthesia, with supplementary succinylcholine. Clinical signs included fever, sweating, hyperventilation, tachycardia, and decreased blood pressure followed by a rapid increase in blood pressure. Biochemical aberrations included hypocalcemia, hyperkalemia, hyperphosphatemia, myoglobinuria, and high creatine phosphokinase and ornithine carbamyl transferase activities. Treatment consisted initially of surface cooling with cold water, alcohol and ice, IV administration of cooled balanced electrolyte solutions and sodium bicarbonate, and removal from the anesthetic and rebreathing circuit. Oxygen was given by endotracheal insufflation. The rectum was then packed with ice, the horse was moved to a recovery raft and pool, and his body was packed in ice. Xylazine and dantrolene were given during recovery from anesthesia. Following recovery, treatment consisted of administration of balanced electrolyte solutions, calcium borogluconate, potassium penicillin, meperidine, and additional dantrolene. Muscle biopsy demonstrated exaggerated contracture responses to halothane and caffeine, confirming a diagnosis of malignant hyperthermia. The horse was returned to training following a routine postsurgical convalescent period.

Complications of umbilical hernias in horses: 13 cases (1972-1986).

Of 147 horses treated for umbilical hernias over a 13.5-year period, 13 horses (8.8%) developed complications in association with umbilical defects. Six horses had intestinal incarceration; the incarceration was reduced manually in 3 horses before admission, resolved without treatment in 2 others, and was surgically reduced in one. Herniorrhaphy was performed on 4 of the 5 horses in which the incarceration did not require surgical reduction, and the fifth was managed conservatively. A horse with a parietal hernia and a horse with intestinal stragulation were treated surgically; in the latter, the involved intestine was resected. These 8 horses recovered. Three horses developed an umbilical abscess and 2 developed an enterocutaneous fistula through their umbilical hernias. Four of these horses responded well to surgery, but one horse with an enterocutaneous fistula died from electrolyte imbalances and peritonitis after an unsuccessful attempt at simple closure. The results of this study confirmed that complications of umbilical hernias are rare in horses; however, when they do develop, they may be one of various forms, some of which are insidious in onset.

Epiglottic augmentation for treatment of dorsal displacement of the soft palate in racehorses: 59 cases (1985-1994).

OBJECTIVE: To determine whether epiglottic augmentation, in conjunction with more traditional surgical methods, would be useful in the treatment of dorsal displacement of the soft palate in racehorses. DESIGN: Retrospective study. ANIMALS: 40 Thoroughbred and 19 Standardbred racehorses. PROCEDURE: Polytetrafluoroethylene paste was injected submucosally on the lingual epiglottic surface of each horse. In addition, sternothyrohyoideus myectomy or sternothyroideus tenectomy and staphylectomy were performed in most horses. RESULTS: Racing performance was improved after surgery in 29 of 40 (73%) Thoroughbreds and 10 of 19 (53%) Standardbreds. Twenty-nine (49%) horses won > or = 1 race after surgery. CLINICAL IMPLICATIONS: Results suggest that epiglottic augmentation, in conjunction with other surgical methods, may be an effective method of treating horses with poor racing performance attributable to dorsal displacement of the soft palate.

Complications during treatment of traumatic disruption of the suspensory apparatus in Thoroughbred horses.

A total of 19 Thoroughbred horses were treated for traumatic disruption of the suspensory apparatus, using either external support of the injured limb, removal of fractured proximal sesamoid bone fragments, metacarpophalangeal arthrodesis, compression screw fixation of the fractured proximal sesamoid bones, application of a cast-brace attached to a transfixation pin inserted through the third metacarpal bone, or combinations thereof. Major complications during the treatment of traumatic disruption of the suspensory apparatus were infection (9 of 19 horses, 47%), large cast sores (10 of 14 treated horses, 71%), laminitis (7 of 19 horses, 37%), and orthopedic implant failure or loosening (4 of 6 treated horses, 67%), which led to euthanasia in 16 of 18 cases with complete follow-up information. Increased pain and lameness signaled the development of such complications. In 7 of 9 cases with infection confirmed by microbiologic culture, the horse had received surgical treatment; in 6 of those 7 cases, the infection involved the surgical site. Postoperative wound infection developed in 4 of 7 cases when the surgery was performed within 18 days of injury. In 3 cases, septic metacarpophalangeal arthritis developed, but it was unrelated to surgical procedures. Implant failure or loosening and infection led to euthanasia in 5 of 6 horses treated by internal fixation to stabilize the metacarpophalangeal joint. Three of 6 attempts to perform metacarpophalangeal arthrodesis by application of a bone plate to the dorsal aspect of the joint resulted in implant failure after 45 to 101 days. Major complications did not develop in 2 horses that were treated successfully.

Pneumocephalus in five horses.

Pneumocephalus is occasionally encountered in horses but poorly described in the literature. The study aimed to describe historical, clinical and diagnostic imaging findings and outcome in horses to increase the awareness and recognition of the condition amongst equine practitioners, allowing appropriate management of these cases. Cases of pneumocephalus from 4 participating institutions were identified and case details extracted from the medical records. Five cases of pneumocephalus were identified. Head trauma with suspected or confirmed fractures of the sinuses was the underlying cause in 4 cases, while the remaining horse was thought to have developed pneumocephalus secondary to sinusitis or trauma. Diagnosis was established radiographically in 4 cases and by computed tomography in a fifth. Gas was noted between the cranial vault and the brain, in the lateral ventricles, in the brain parenchyma and within the cranial cervical canal. The gas accumulation resolved gradually in all horses and did not appear independently to result in neurological compromise. Long-term outcome was available for 3 cases, 2 of which were ultimately subjected to euthanasia due to neurological deficits. Pneumocephalus is a possible consequence of head trauma or sinusitis in horses; although the finding is frequently incidental, it has the potential to develop into a life-threatening complication. Imaging the complete skull and cranial cervical spine is important to allow identification and appropriate management of these cases. The use of computed tomography enables easier identification and localisation of the gas accumulation within the central nervous system.

Effects of a novel hydrogel on equine bone healing: a pilot study.

OBJECTIVE: To examine the efficacy and biocompatibility of a thiolated gelatin-thiolated carboxymethyl hyaluronan (CMHA-SGX) sponge as an osteoconductive device in an equine second and fourth metacarpal bone defect model. METHODS: Seven millimetre segmental ostectomies were created bilaterally in the second and fourth metacarpal bones of four horses. The left and right metacarpal defects were randomly assigned to (1) be filled with a CMHA-SGX sponge (treated) or (2) were left unfilled (control). The duration of the study was nine weeks. Bone healing was evaluated using serial radiology, as well as histologically and histomorphometrically. Data were analyzed using an analysis of variance (ANOVA). The level of significance was p<0.05. RESULTS: Serial radiographic evaluation revealed improved healing in the treated compared to the control defects at weeks eight and nine (p = 0.02). This finding was not corroborated histologically. Histomorphometry did not reveal any significant differences in healing between experimental groups. The CMHA-SGX sponge did not inhibit bone formation, induce local inflammation or lead to surgical site infection. CLINICAL SIGNIFICANCE: While further optimization to improve osteoconductive properties should be considered, the CMHA-SGX sponge appears to be a biocompatible orthopaedic implant and its use as a carrier for osteogenic proteins warrants further investigation.