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Purpose: Brain metastases can occur in up to 50% of patients with metastatic HER2-positive breast cancer. Because patients with active brain metastases were excluded from previous pivotal clinical trials, the central nervous system (CNS) activity of the antibody–drug conjugate trastuzumab deruxtecan (T-DXd) is not well characterized. Experimental Design: We studied how T-DXd affects growth and overall survival in orthotopic patient-derived xenografts (PDX) of HER2-positive and HER2-low breast cancer brain metastases (BCBM). Separately, we evaluated the effects of T-DXd in a retrospective cohort study of 17 patients with stable or active brain metastases. Results: T-DXd inhibited tumor growth and prolonged survival in orthotopic PDX models of HER2-positive (IHC 3+) and HER2-low (IHC 2+/FISH ratio < 2) BCBMs. T-DXd reduced tumor size and prolonged survival in a T-DM1–resistant HER2-positive BCBM PDX model. In a retrospective multi-institutional cohort study of 17 patients with predominantly HER2-positive BCBMs, the CNS objective response rate (ORR) was 73% (11/15) while extracranial response rate was 45% (5/11). In the subset of patients with untreated or progressive BCBM at baseline, the CNS ORR was 70% (7/10). The median time on treatment with T-DXd was 8.9 (1.3–16.2) months, with 42% (7/17) remaining on treatment at data cutoff. Conclusions: T-DXd demonstrates evidence of CNS activity in HER2-positive and HER2-low PDX models of BCBM and preliminary evidence of clinical efficacy in a multi-institution case series of patients with BCBM. Prospective clinical trials to further evaluate CNS activity of T-DXd in patients with active brain metastases are warranted. See related commentary by Soffietti and Pellerino, p. 8

Purpose:Brain metastases can occur in up to 50% of patients with metastatic HER2-positive breast cancer. Because patients with active brain metastases were excluded from previous pivotal clinical trials, the central nervous system (CNS) activity of the antibody–drug conjugate trastuzumab deruxtecan (T-DXd) is not well characterized.Experimental Design:We studied how T-DXd affects growth and overall survival in orthotopic patient-derived xenografts (PDX) of HER2-positive and HER2-low breast cancer brain metastases (BCBM). Separately, we evaluated the effects of T-DXd in a retrospective cohort study of 17 patients with stable or active brain metastases.Results:T-DXd inhibited tumor growth and prolonged survival in orthotopic PDX models of HER2-positive (IHC 3+) and HER2-low (IHC 2+/FISH ratio < 2) BCBMs. T-DXd reduced tumor size and prolonged survival in a T-DM1–resistant HER2-positive BCBM PDX model. In a retrospective multi-institutional cohort study of 17 patients with predominantly HER2-positive BCBMs, the CNS objective response rate (ORR) was 73% (11/15) while extracranial response rate was 45% (5/11). In the subset of patients with untreated or progressive BCBM at baseline, the CNS ORR was 70% (7/10). The median time on treatment with T-DXd was 8.9 (1.3–16.2) months, with 42% (7/17) remaining on treatment at data cutoff.Conclusions:T-DXd demonstrates evidence of CNS activity in HER2-positive and HER2-low PDX models of BCBM and preliminary evidence of clinical efficacy in a multi-institution case series of patients with BCBM. Prospective clinical trials to further evaluate CNS activity of T-DXd in patients with active brain metastases are warranted.See related commentary by Soffietti and Pellerino, p. 8

Figures S1 and S2

Background It is unclear whether diabetes and glycemic control affects the outcomes of breast cancer, especially among those with metastatic disease. This study aims to determine the impact of diabetes and hyperglycemia on cancer progression and mortality in individuals with metastatic breast cancer (MBC). Methods Patients with a diagnosis of MBC between 2010 and 2021 were identified using the MBC database at 2 academic institutions. We evaluated the effects of diabetes and glycemic control on overall survival (OS) and time to next treatment (TTNT). Results We compared 244 patients with diabetes (median age 57.6 years) to 244 patients without diabetes (matched for age, sex, ethnicity, and receptor subtype). OS at 5 years [diabetes: 54% (95% CI 47-62%) vs controls: 56% (95% CI 49-63%), P = 0.65] and TTNT at 1 year [diabetes: 43% (95% CI 36-50%) vs controls: 44% (95% CI 36-51%), P = 0.33] were similar between groups. A subgroup analysis comparing those with good glycemic control and those with poor glycemic control among patients with specific receptor subtype profiles showed no differences in OS at 5 years or TTNT at 1 year. In an 8-year landmark subgroup analysis, there was worse OS among individuals with diabetes compared to controls, and OS was found to be better among those with good glycemic control compared to those with poor control. Conclusions Diabetes was not associated with increased mortality in individuals with MBC at 5 years. However, diabetes and hyperglycemia were associated with worse OS among a cohort of longer-term survivors. These findings suggest that individualized diabetes and glycemic goals should be considered in patients with MBC.

Background: Although immune checkpoint inhibitors (ICI) are most widely used in PD-L1 positive, metastatic triple-negative breast cancer (TNBC), the clinical utility of ICI in other tumor subtypes and patient populations is not well defined. In this study, we measured infiltration of multiple immune cell populations in primary and metastatic HR+ tumors to infer potential benefit from ICI. We investigated how risk stratifying features such as pathologic grade and Oncotype Dx scores affect immune cell infiltration across subtypes. Lastly, we compared immune cell infiltration in breast tumors from younger versus older women, as our pre-clinical studies demonstrated differential enrichment of immune response pathways by age and association with improved response to ICI. Methods: ImmunoProfile, a multiplexed immunofluorescence (IF) assay, was performed on FFPE tissue from 167 archived breast cancer tumors from the Dana-Farber Cancer Institute (HR+/HER2- n=124, HER2+ n=21, TNBC n=22). Unmatched samples were obtained from either primary breast (HR+/HER2- n=104, HER2+ n=13, TNBC n=15) or metastatic core biopsies. IF staining signal was measured for T cell markers CD8 and co-localized CD8/PD-1, T regulatory cell marker FOXP3, PD-L1 and PD1 expression on tumor cells as well as PD-L1 expression on inflammatory cells. Inflammatory, tumoral (TPS) and combined inflammatory and tumor (CPS) pathologic PD-L1 scores were calculated. PD-L1 staining was also assessed by IHC (E1L3N) staining and compared to multiplexed IF stains. IF staining signal (Vectra analyzed, log2+1 transformed) was analyzed against clinical and pathologic variables, including tumor subtype, primary versus metastatic status, pathologic grade, and Oncotype Dx score, using non-parametric statistical comparisons. Given the decline in adaptive immunity that occurs with normal physiological aging and our own group’s previous reports, we also explored whether immune cell staining signal associated with age. Results: TPS, CPS, and inflammatory PD-L1 scores strongly correlated between IHC and IF staining methods (Spearman r = 0.660, 0.683, 0.638 respectively, p < 2.2e-16). When HR+/HER2- (n=124), HER2+ (n=21) and TNBC (n=22) cases were analyzed separately, there was no statistical difference between unmatched primary and metastatic cases in any of the analyzed immune cell population stains for any subtype. When all primary cases were analyzed, TNBC showed higher CD8, CD8/PD1, PD1 (all p < 0.05), and FOXP3 staining as well as IF CPS, TPS and inflammatory PD-L1 scores (all p < 0.005) than HR+ cases. HR+/HER2- cases from women < age 65 (n=56) showed higher intra-tumoral CD8 (p=0.039), PD1 (p=0.043), and CD8/PD1 (p=0.022) than cases from women > age 65 (n=38). There was a statistically significant step-wise increase in IF staining of all immune cell populations with increasing pathologic grade (I, II, III) as well as Oncotype Dx scores (< 11, 11-25, >25) among primary HR+/HER2- cases (n=74). Conclusions: Strong correlations between PD-L1 scores based on IF and IHC methods suggest clinical utility for multiplexed IF immune cell staining in therapeutic planning. ImmunoProfile demonstrated the previously known pattern that primary TNBC contains higher immune cell infiltration than HR+ or HER2+ tumors. Women < age 65 with HR+ cancer showed higher levels of CD8 and PD1 stained populations, suggesting there may be greater utility of ICI in younger patients. Lastly, the association of higher pathologic grade and Oncotype Dx score with increasing immune cell infiltration supports exploration of ICI in biologically unfavorable primary HR+ cancers. Citation Format: Tess A. O'Meara, Tanya E. Keenan, Adrienne G. Waks, Kristen D. Felt, Bijaya Sharma, Scott Rodig, Melissa Hughes, Nancy U. Lin, Judith Agudo, Jennifer L. Guerriero, Sandra S. McAllister, Elizabeth Mittendorf, Sara Tolaney. Multiplexed immunofluorescence staining of intra-tumoral immune cell populations and associations with immunohistochemical, clinical, and pathologic variables in breast cancer [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P1-04-05.

Kaplan-Meier Curves for Progression-Free Survival and Overall Survival by high TMB {greater than or equal to}7 mutations/Mb and PTEN alterations in Chemotherapy-Treated mTNBC. (A) Progression-free survival and (C) overall survival by tumor mutational burden

Purpose:Few patients with metastatic triple-negative breast cancer (mTNBC) benefit from immune checkpoint inhibitors (ICI). On the basis of immunotherapy response correlates in other cancers, we evaluated whether high tumor mutational burden (TMB) ≥10 nonsynonymous mutations/megabase and PTEN alterations, defined as nonsynonymous mutations or 1 or 2 copy deletions, were associated with clinical benefit to anti-PD-1/L1 therapy in mTNBC.Experimental Design:We identified patients with mTNBC, who consented to targeted DNA sequencing and were treated with ICIs on clinical trials between April 2014 and January 2019 at Dana-Farber Cancer Institute (Boston, MA). Objective response rate (ORR), progression-free survival (PFS), and overall survival (OS) were correlated with tumor genomic features.Results:Sixty-two women received anti-PD-1/L1 inhibitors alone (23%) or combined with targeted therapy (19%) or chemotherapy (58%). High TMB (18%) was associated with significantly longer PFS (12.5 vs. 3.7 months; P = 0.04), while PTEN alterations (29%) were associated with significantly lower ORR (6% vs. 48%; P = 0.01), shorter PFS (2.3 vs. 6.1 months; P = 0.01), and shorter OS (9.7 vs. 20.5 months; P = 0.02). Multivariate analyses confirmed that these associations were independent of performance status, prior lines of therapy, therapy regimen, and visceral metastases. The survival associations were additionally independent of PD-L1 in patients with known PD-L1 and were not found in mTNBC cohorts treated with chemotherapy (n = 90) and non-ICI regimens (n = 169).Conclusions:Among patients with mTNBC treated with anti-PD-1/L1 therapies, high TMB and PTEN alterations were associated with longer and shorter survival, respectively. These observations warrant validation in larger datasets.

Extended methods and results, including e-references

In monogamous mating systems, pair mates frequently share similar characteristics, such as size or ornamentation, a pattern suggesting mutual mate choice. When pairs persist across multiple seasons, preferences for familiar pair mates can either reinforce or disrupt patterns of assortative pairing. Snapping shrimp (Alpheus angulosus) are socially monogamous shrimp that can be found in size-assortative pairs year-round; both sexes could benefit from pairing with larger individuals. To determine the role of pair mate preferences for either size or established pair mates in size-assortative pairing, we performed simultaneous choice trials to test whether (1) either or both sexes prefer larger pair mates, (2) females prefer males with larger claws, or (3) either or both sexes prefer current pair mates over size-matched unfamiliar potential mates. Males, but not females, preferred larger potential pair mates in both the reproductive and nonreproductive seasons; males also showed more mate-searching behaviour, switching more frequently between choice options than females. Females, but not males, preferred their current pair mate over a novel potential pair mate; females also engaged in more frequent switching between pair mates and novel males, but only during the nonreproductive season. Thus, while both males and females exhibit preferences in pairing, size-assortative pairing in snapping shrimp does not result from mutual mate choice for size, but rather preferences for size in one sex, with size-assortative pairing reinforced by preferences for established pair mates in the other. Furthermore, pairing behaviour may be dynamic, shifting across seasons even in a year-round socially monogamous species.

Background While a well-established relationship between diabetes and breast cancer exists, it remains unclear how diabetes impacts breast cancer outcomes. This study aims to determine the impact/association of diabetes and hyperglycemia on cancer progression and mortality in individuals with metastatic breast cancer. Methods Patients with metastatic breast cancer between 2010-2021 were identified using the metastatic breast cancer database at a single academic center. The Research Patient Data Registry, a data warehouse of all patients receiving care in the hospital network was then used to determine the diabetes status of each participant. A participant was categorized as having diabetes if they fulfilled pre-specified criteria (international classification of diseases (ICD) 9 or 10 code for diabetes, or a hemaglobin A1c ≥6.5%, or a random blood glucose (RBG) ≥200 mg/dL and documented use of glucose-lowering agents), while controls were defined as individuals who did not fulfill the previous criteria. We evaluated the effects of diabetes on 1) overall survival (OS) and 2) the interval from initiation of first-line metastatic therapy to initiation of a second-line regimen due to documented progression of disease (a surrogate measure of progression-free survival). The impact of glycemic control on OS and duration of first-line therapy was also assessed. Results We compared 244 patients with diabetes (median age 57.6 years) to 244 patients without diabetes, who were matched for age, gender, ethnicity and hormone-receptor (HR) status. OS at 5 years (diabetes: 54%, CI; 47-62% vs. controls: 56%, CI; 49-63%, p=0.65) and freedom from initiating a second-line regimen at 2 years (diabetes: 43%, CI; 36-50% vs. controls: 44%, CI; 36-51%, p=0.33) were not statistically different between groups. However, in an 8-year landmark subgroup analysis, OS was better amongst controls than those with diabetes (diabetes [n=27]: 67%, CI; 48-86% vs. Controls [n=28]: 87%, CI; 73-100%, p=0.047at 10 years). Poor glycemic control (median RBG >180 mg/dL or median HbA1c >7%) was not associated with increased mortality at 5 years but was associated with a trend towards worse freedom from initiating a second-line regimen at 2 years when compared to good glycemic control. In a sub-group analysis based on HR status, there were no differences in OS at 5 years, or freedom from initiating a second-line regimen at 2 years between the two glycemic groups (≤180 mg/dL vs >180 mg/dL on two or more occasions in a month) for any of the HR subgroups. Conclusions These data provide some reassurance that hyperglycemia may not be a major contributor to overall mortality in the first 5 years, in most individuals with metastatic breast cancer. However, amongst longer-term survivors, diabetes was associated with worse survival, suggesting that individualized diabetes and glycemic goals should, therefore, be considered in patients with metastatic breast cancer. Presentation: Sunday, June 12, 2022 12:30 p.m. - 2:30 p.m., Monday, June 13, 2022 12:51 p.m. - 12:56 p.m.

We sought to understand the attitudes of individuals with abnormal breast imaging findings prompting a diagnostic breast biopsy toward donation of blood, excised tissue, or percutaneous biospecimens for research, and to understand medical oncologists' attitudes toward research biospecimen collection in this population.We included individuals who presented to a single academic medical center for a clinically indicated, image-guided, percutaneous breast biopsy. We administered a survey prior to knowledge of biopsy results to assess willingness to consider, entirely for research purposes, donating blood or excess excised breast tissue, or having additional biospecimens (AB) obtained during a clinically indicated percutaneous biopsy. We also surveyed breast medical oncologists from National Cancer Institute-designated cancer centers to assess attitudes toward approaching patients for biospecimen research.Overall, 53/63 patients responded to the survey; 70% would consider donating blood, 85% would consider donating excess excised breast tissue, and 32% would consider having AB obtained during a clinically indicated biopsy. Main motivating factors for considering AB included contributing to scientific knowledge and return of study or biopsy results, whereas anxiety and the potential discomfort were the main dissuading factors. Among 191 medical oncologists, most were very comfortable (59.2%), or somewhat comfortable (32.5%) asking patients to have AB obtained during a clinically indicated breast biopsy. Medical oncologists reported hesitancy to refer a patient for AB due to potential pain/discomfort, and other procedure risks.Only one-third of individuals with breast imaging findings would consider consenting to AB during a diagnostic biopsy, whereas most were open to donating blood or excess excised breast tissue. Most medical oncologists would be comfortable asking patients to have AB obtained during the biopsy. Understanding patients' and oncologists' baseline attitudes may inform the design and approach to breast biospecimen-based research.

Purpose: Up to half of patients (pts) with HER2+ metastatic breast cancer (MBC) will develop BCBM. Trastuzumab deruxtecan (T-DXd) is an antibody-drug conjugate (ADC) with demonstrated efficacy in previously treated pts with HER2+ MBC. However, few pts with stable/treated brain metastases, and no pts with active (untreated or progressive) brain metastases, were included in completed clinical trials of T-DXd. Thus, central nervous system (CNS) efficacy of T-DXd is not well-characterized. Methods: We tested T-DXd in orthotopic pt BCBM-derived xenograft (PDX) models of HER2+, and HER2-low, BCBMs. To further validate T-DXd single agent CNS activity, we described clinical outcomes of T-DXd in a multi-institutional retrospective cohort of 16 patients with BCBM. Consecutive pts who initiated T-DXd between 1 Jan 2020 - 1 Nov 2020 (Duke) or 1 Jan 2020 - 15 June 2020 (Dana-Farber Cancer Institute) were included. Data cut-off date was 31 Dec 2020. CNS response was measured by via central radiology review at each participating institution. Up to 5 CNS target lesions were included. CNS partial response (PR) required >30% reduction in sum of CNS target lesions. Results: Treatment with 10 mg/kg T-DXd significantly prolonged survival in HER2+ BCBM PDX models DFBM-354 (67 vs 154 days, p=0.0018) and DFBM-355 (78 vs 156 days, p=0.0067) vs. vehicle control. We then tested TDX-d in a HER2 low BCBM PDX model (IHC 2+/FISH ratio Table 1.Best CNS Response of study cohortOverall population (n=16)Pts with progressive or untreated CNS disease at baseline (N=9)Pts with stable/treated CNS disease at baseline (N=7)Complete response (CR)0 (0)00 (0)Partial response (PR)10 (63)6 (67)#4 (57)+Stable disease (SD)2 (13)2 (22)0 (0)Progressive disease (PD)1 (6)0 (0)1 (14)No measurable CNS disease at baseline2 (13)0 (0)2 (29)Lost to follow-up1 (6)1 (11)0 (0).#Among these 6 pts, interval between most recent radiation and T-DXd initiation was 14.3 months, 12.4 months, 17 months, 18.2 months, and 8.2 months (one pt was radiation-naïve). +Among these 4 pts, interval between most recent radiation and T-DXd initiation was 19.1 months, 15.1 months, 1.5 months, and 16.9 months, respectively. Citation Format: Sheheryar Kabraji, Jing Ni, Sarah Sammons, Amanda ED Van Swearingen, Yanzhi Wang, Alyssa M Pereslete, Liangge Hsu, Chris Lascola, Heather Moore, Melissa Hughes, Akshara S Raghavendra, Maria Gule-Monroe, Rashmi K Murthy, Eric P Winer, Carey K Anders, Jean J Zhao, Nancy U Lin. Preclinical and clinical efficacy of trastuzumab deruxtecan in breast cancer brain metastases (BCBM) [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr PD4-05.

Sexually dimorphic weaponry often results from intrasexual selection, and weapon size can vary seasonally when costs of bearing the weapon exceed the benefits outside of the reproductive season. Weapons can also be favored in competition over nonreproductive resources such as food or shelter, and if such nonreproductive competition occurs year‐round, weapons may be less likely to vary seasonally. In snapping shrimp (Alpheus angulosus), both sexes have an enlarged snapping claw (a potentially deadly weapon), and males of many species have larger claws than females, although females are more aggressive. This contrasting sexual dimorphism (larger weaponry in males, higher aggression in females) raises the question of whether weaponry and aggression are favored by the same mechanisms in males and females. We used field data to determine whether either sex shows seasonal variation in claw size such as described above. We found sexual dimorphism increased during the reproductive season due to opposing changes in both male and female claw size. Males had larger claws during the reproductive season than during the nonreproductive season, a pattern consistent with sexual selection. Females, however, had larger claws during the nonreproductive season than during the reproductive season—a previously unknown pattern of variation in weapon size. The observed changes in female weapon size suggest a trade‐off between claw growth and reproduction in the reproductive season, with investment in claw growth primarily in the nonreproductive season. Sexually dimorphic weaponry in snapping shrimp, then, varies seasonally due to sex differences in seasonal patterns of investment in claw growth, suggesting claws may be advantageous for both sexes but in different contexts. Thus, understanding sexual dimorphisms through the lens of one sex yields an incomplete understanding of the factors favoring their evolution.

Highlights • Breast cystosarcomaphyllodes tumors are rare and can be benign or malignant. • These tumors can harbor carcinomas, although the incidence is extremely rare. • Although challenging, the detection of a carcinoma component in phyllodes tumor is important, as it can dictate the need for lymph node sampling and possible adjuvant therapies such as radiation and systemic management., Introduction Breast cystosarcoma phyllodes tumors are rare and can be benign or malignant. All sub-divisions of phyllodes tumor—benign, borderline and malignant, can harbor carcinomas, although the incidence is extremely rare. Methods: We present two nonconsecutive cases of coexisting ductal carcinoma in situ (DCIS) and phyllodes breast tumors in young patients. Methods & Case Presentation Retrospective review of two patient’s medical record was performed. Case 1 30-year-old female underwent excisional biopsy for 3.48 cm mass found on ultrasound. Pathology revealed malignant phyllodes tumor with positive margin. On re-excision, patient was found to have 1.5 cm area of ductal carcinoma in situ (DCIS) with positive margin. Patient then underwent re-reexicision of DCIS with negative margin. Patient underwent chemotherapy and tamoxifen for three years without evidence of disease. Case 2 30-year-old female presented with 1.3 cm lesion found on ultrasound which core needle biopsy revealed a fibroepithelial tumor. Patient subsequently underwent excision biopsy which found 1.5 cm benign phyllodes tumor and 3.5 mm DCIS within the phyllodes tumor with negative margins. Patient declined additional chemotherapy or hormonal therapy and is currently considering mastectomy. Conclusion Phyllodes tumors are rare and ones with a coexisting carcinoma are even less frequently encountered. The treatment plan can change upon diagnosis of the carcinoma via the pathology. Treatment should be guided by the type and stage of carcinoma detected which may include additional surgical resection and lymph node sampling.

Behavior courses face numerous challenges when moving to an online environment, as has been made necessary by the COVID‐19 pandemic. These challenges occur largely because behavior courses, like most organismal biology courses, often stress experiential learning through laboratories that involve live animals, as well as a lecture component that emphasizes formative assessment, discussion, and critical thinking. Although online behavior courses may be remote, they can still be interactive and social, and designed with inclusive pedagogy. Here, we discuss some of the key decisions that instructors should consider, provide recommendations, and point out new opportunities for student learning that stem directly from the move to online instruction. Specific topics include challenges related to generating an inclusive and engaging online learning environment, synchronous versus asynchronous formats, assignments that enhance student learning, testing format and execution, grade schemes, design of laboratory experiences including opportunities for community science, design of synthetic student projects, and workload balance for students and instructors. We designed this primer both for animal behavior instructors who need to quickly transition to online teaching in the midst of a pandemic, and for those facing such transitions in upcoming terms. Much of the manuscript's content should also be of general interest and value to instructors from all areas of organismal biology who are attempting to quickly transition to online teaching., Teaching behavior online presents both challenges and opportunities. In this comprehensive guide, we offer suggestions for the design and implementation of online behavior courses. For all aspects of these courses, including lectures, discussions and labs, we emphasize a backwards‐design approach and the importance of flexibility, inclusivity, equity, and accessibility. We also offer recommendations for delivery mode, specific course components, and resources for additional support.

Face-to-face classes in animal behavior often stress experiential learning through laboratories that involve observation of live animals, as well as a lecture component that emphasizes formative assessment, discussion and critical thinking. As a result, behavior courses face unique challenges when moving to an online environment, as has been made necessary at many institutions due to the COVID-19 pandemic. Although online behavior courses may be remote, they can still be interactive and social, and designed with inclusive pedagogy. Here we discuss some of the key decisions that instructors should consider, provide recommendations, and point out new opportunities for student learning that stem directly from the move to online instruction. Specific topics include challenges related to generating an inclusive and engaging online learning environment, synchronous versus asynchronous formats, assignments that enhance student learning, testing format and execution, grade schemes, design of laboratory experiences including opportunities for Community Science, design of synthetic student projects, and workload balance for students and instructors.

We aimed to describe our experience with metaplastic breast carcinoma (MBC), evaluate its clinical outcome compared with triple-negative breast cancer (TNBC), and provide a through and comprehensive review of the literature to date.We reviewed MBC cases (n = 46) from our institution. The following variables were recorded: tumor histologic subtype, Nottingham grade, tumor size, lymph node status, Tumor, Node, Metastases stage, biomarkers profile, patient's age and race, therapy modality (chemotherapy and radiation), and survival (disease-free survival [DFS] and overall survival [OS]). The clinical and pathological data for TNBC (n = 508) cases were extracted from the breast cancer database. To compare the survival between MBC and TNBC, a subgroup of MBC cases (n = 40) were matched with TNBC cases (n = 40) on the basis of known prognostic confounders.There were 17 of 46 (37%) cases with mesenchymal differentiation, 12 (26.1%) squamous cell carcinoma, 14 (30.4%) spindle cell carcinoma, and 3 (6.5%) mixed type. MBC presented at a more advanced stage than TNBC (P = .014) and was more likely to recur (34% vs. 15.5%; P = .004). More MBC patients died from disease than TNBC (29% vs. 16%; P = .05). In the multivariate analysis, MBC had approximately twice the risk of local recurrence than TNBC (95% confidence interval, 1.01-3.83; P = .05). MBC patients had worse DFS and OS than the matched TNBC patients (P .001 and P = .033, respectively). A review of the literature comparing MBC versus TNBC is presented.Our results suggest that MBC is clinically more aggressive than TNBC. Further studies might help delineate the differences between these 2 entities.

The green porcelain crab, Petrolisthes armatus, is a common invasive species on inter-tidal oyster reefs in the South Atlantic Bight whose behavior is largely unknown. We assessed the effects of the presence of opposite-sex conspecifics on adult crab spacing at a low density to infer potential mate acquisition behaviors. Adult crabs held in all-male, all-female, and mixed-sex treatments were observed twice daily over five days to determine nearest-neighbor (NN) distances and relative activity. Males in the mixed-sex treatment with the shortest NN distances (NN1) were most commonly (1) closest to females, and, when compared to NN1 males in the all-male treatment, were (2) closer to their NN and (3) had lower activity levels. Female movement did not differ between treatments. Percent ovigerous females at the end of trials were higher in the mixed-sex treatment, reflecting mating activity, which when observed, involved inter-molt females. Mate guarding, rather than pure-searching behavior, was the ma...

PURPOSE: Few patients with metastatic triple-negative breast cancer (mTNBC) benefit from immune checkpoint inhibitors (ICIs). Based on immunotherapy response correlates in other cancers, we evaluated whether high tumor mutational burden (TMB) ≥10 nonsynonymous mutations/megabase and PTEN alterations, defined as nonsynonymous mutations or 1 or 2 copy deletions, were associated with clinical benefit to anti-PD-1/ L1 therapy in mTNBC. EXPERIMENTAL DESIGN: We identified patients with mTNBC, who consented to targeted DNA sequencing and were treated with ICIs on clinical trials between April 2014 and January 2019 at Dana-Farber Cancer Institute (Boston, MA). Objective response rate (ORR), progression-free survival (PFS), and overall survival (OS) were correlated with tumor genomic features. RESULTS: Sixty-two women received anti-PD-1/L1 inhibitors alone (23%) or combined with targeted therapy (19%) or chemotherapy (58%). High TMB (18%) was associated with significantly longer PFS (12.5 vs. 3.7 months; p=0.04); while PTEN alterations (29%) were associated with significantly lower ORR (6% vs. 48%; p=0.01), shorter PFS (2.3 vs. 6.1 months; p=0.01), and shorter OS (9.7 vs. 20.5 months; p=0.02). Multivariate analyses confirmed that these associations were independent of performance status, prior lines of therapy, therapy regimen, and visceral metastases. The survival associations were additionally independent of PD-L1 in patients with known PD-L1 and were not found in mTNBC cohorts treated with chemotherapy (n = 90) and non-ICI regimens (n = 169). CONCLUSIONS: Among patients with mTNBC treated with anti-PD-1/L1 therapies, high TMB and PTEN alterations were associated with longer and shorter survival, respectively. These observations warrant validation in larger datasets.

Territoriality is a special case of resource defense, in which space is actively defended for exclusive use. As active defense is likely to be costly, territoriality is expected only when the benefits of exclusivity outweigh these costs. In most territorial species of noncrustacean taxa, the defended space includes resources critical for reproduction or food. These resources are not only critical for reproductive success, but also are vulnerable to “looting”, that is, the value of these resources may be reduced through short-term intrusions, even without loss of ownership, thus providing an advantage for active defense of exclusive space. Many crustaceans defend space, particularly burrows or other shelters that are refuges from predation or environmental stressors. While protection is obviously a critical resource, it is not a resource that necessarily requires exclusivity; indeed, many crustaceans that depend upon shelters for protection do not defend them for exclusive use. Nonetheless, many crustacean taxa aggressively defend exclusive access to their shelters. Crustaceans, then, may be especially suitable for testing alternative hypotheses of territoriality, including the potential benefits of interindividual spacing rather than defense of space per se. It is also worth considering a null hypothesis for territoriality: aggressive defense of space in crustaceans may be an artifact of relatively sedentary species with high intraspecific aggression favored in other contexts, rather than aggression favored for defense of particular resources. In addition to these questions, much remains to be learned about territorial behaviors in crustaceans. Most notably, the boundaries of defended space are unknown in many taxa. Understanding the boundaries of defended space is important for understanding the ecological consequences of territoriality, as well as aspects of territory acquisition and the roles of neighbor relationships and territorial advertisement signals in territory defense. Many crustacean territories appear to differ from those described for other animals, especially terrestrial species; it is not clear, however, whether these differences are due to differences in function or habitat, or rather result from our incomplete knowledge of crustacean territoriality.

The snapping shrimp,Alpheus angulosusMcClure, 2002, is a small crustacean with bilaterally asymmetric claws that serve distinct behavioural and sensory functions. If the large claw is lost, the organism switches handedness, transforming its small pincer claw into a large snapping claw while simultaneously developing a small claw on the contralateral side. To better understand the mechanisms required to adapt to this radical change in body composition, we examined developmental plasticity by tracing changes in sensory setae distribution on the claws throughout transformation. We observed only two broad types of setae, simple and plumose. Quantitative analysis across molt stages revealed significant alterations in setae composition and numbers that occurred primarily on the edge of the propodus, where the most drastic morphological changes also occur. These results suggest that previous developmental mechanisms are re-engaged to support the proliferation and differentiation of new setae during transformation.

In his introduction to Michael Anderson and Colleen Roche’s, ‘The State of the Art’ Robin Pascoe depicts this collection as a ‘powerful frame through which to make sense of the unfolding turbulent ...

This review was developed at an investigative workshop, “Analyzing Animal Vocal Communication Sequences” that took place on October 21–23 2013 in Knoxville, Tennessee, sponsored by the National Institute for Mathematical and Biological Synthesis (NIMBioS). NIMBioS is an Institute sponsored by the National Science Foundation, the U.S. Department of Homeland Security, and the U.S. Department of Agriculture through NSF Awards #EF-0832858 and #DBI-1300426, with additional support from The University of Tennessee, Knoxville. In addition to the authors, Vincent Janik participated in the workshop. D.T.B.’s research is currently supported by NSF DEB-1119660. M.A.B.’s research is currently supported by NSF IOS-0842759 and NIH R01DC009582. M.A.R.’s research is supported by ONR N0001411IP20086 and NOPP (ONR/BOEM) N00014-11-1-0697. S.L.DeR.’s research is supported by the U.S. Office of Naval Research. R.F.-i-C.’s research was supported by the grant BASMATI (TIN2011-27479-C04-03) from the Spanish Ministry of Science and Innovation. E.C.G.’s research is currently supported by a National Research Council postdoctoral fellowship. E.E.V.’s research is supported by CONACYT, Mexico, award number I010/214/2012.

Sexual dimorphisms in weaponry and aggression are common in species in which one sex (usually males) competes for access to mates or resources necessary for reproduction – sexually dimorphic weaponry and aggression, in other words, are frequently the result of intrasexual selection. In snapping shrimp, the major chela (snapping claw) can be a deadly weapon, and males of many species have larger chelae than females, a pattern readily interpreted as resulting from intrasexual selection. Thus, males might be expected to show more sex-specific aggression than females, and be more aggressive overall. We tested these predictions in two species of snapping shrimp in a territorial defense context. Neither of these predictions was supported: in both species, females, but not males, engaged in sex-specific aggression and females were more aggressive than males overall. These contrasting sexual dimorphisms – larger weaponry in males but higher aggression in females – highlight the importance of considering the function of weaponry and aggression in contexts other than direct competitions over mates. In addition, species differences in the degree of sexual dimorphism in chela size were due to differences in female, not male, chela size, and the species with greater sexual dimorphism in weaponry was significantly less aggressive overall; also, while paired and solitary males did not differ in residual chela size, for the species with greater sexual dimorphism, females carrying embryos had smaller residual chela sizes. These results suggest that understanding the sexual dimorphisms in weaponry and aggression in snapping shrimp requires understanding the relative costs and benefits of both in females as well as males.

The ability to regenerate lost tissues, organs or whole body parts is widespread across animal taxa; in some animals, regeneration includes transforming a remaining structure to replace the one that was lost. The transformation of one limb into another involves considerable plasticity in morphology, physiology and behavior, and snapping shrimp offer excellent opportunities for studying this process. We examined the changes required for the transformation of the small pincer to a mature snapping claw in Alpheus angulosus. First molt claws differ from mature claws in overall shape as well as in morphology related to snapping function; nonetheless, shrimp with first molt claws do produce snaps. While most shape variables of second molt claws do not differ significantly from mature claws, the plunger (structure required for snap production) does not reach mature size until the third molt for females, or later for males. Thus, the pincer claw can be transformed into a functional snapping claw in one molt, alth...

While the relationship between aggression and resource value in competitions for discrete resources is well established, the relationship between aggression and territory quality is less well understood. Territoriality imposes spatial structure on social interactions: if spatial heterogeneity in habitat quality leads to clustering of high- and low-quality territories, and if consistently more aggressive individuals are more likely to obtain higher-quality territories, the resulting clusters of territory holders of similar aggression are likely to influence a wide array of social dynamics in territorial species, particularly when aggressive behaviour is consistently associated with variation in other behaviours, such as boldness. Using 8 years of historical nesting data in a northeastern population of song sparrows, we here determine (1) whether historical nesting success (defined both as clutch size and survival) is spatially clustered; and (2) whether aggression or boldness in first-year territorial males is correlated with the historical clutch size or nest survival of each male's territory. We found significant clustering of both predation risk and large clutch sizes, even when controlling for the identities of historical territory owners. The aggressive behaviour of new territorial males was correlated with historical clutch size, but not with historical nest survival; in contrast, boldness was not correlated with either historical clutch size or nest survival. Thus, spatial variation in habitat quality is correlated with territorial aggression, but territory holders may either prioritize or be unable to assess all variables contributing to territory quality. In addition, a boldness-aggression syndrome does not necessarily result in both behaviours covarying with territory quality.

Alpheus angulosus McClure, 2002 is one of several species of snapping shrimp that live along the east coast of the United States and belong to the edwardsii group of Alpheus. The genus Alpheus presents with bilateral asymmetry in their chelipeds, specifically a large snapper and a smaller pincer. This is an extreme example of the asymmetry found in many other crustaceans. A significant amount of work has been done on the adult behavior, physiology, and transformation/regeneration of the two claws, but less is known about the early development of the nervous system that underlies this asymmetry. The work reported here begins to establish an atlas of embryonic development in this species staged by using both eye index and percent development connected to yolk depletion during embryogenesis. This represents the first step toward a more comprehensive understanding of embryonic development that can be used to address future neuro-developmental questions regarding limb asymmetry and plasticity.

Learned aspects of song have been hypothesized to signal cognitive ability in songbirds. We tested this hypothesis in hand-reared song sparrows (Melospiza melodia) that were tutored with playback of adult songs during the critical period for song learning. The songs developed by the 19 male subjects were compared to the model songs to produce two measures of song learning: the proportion of notes copied from models and the average spectrogram cross-correlation between copied notes and model notes. Song repertoire size, which reflects song complexity, was also measured. At 1 year of age, subjects were given a battery of five cognitive tests that measured speed of learning in the context of a novel foraging task, color association, color reversal, detour-reaching, and spatial learning. Bivariate correlations between the three song measures and the five cognitive measures revealed no significant associations. As in other studies of avian cognition, different cognitive measures were for the most part not correlated with each other, and this result remained true when 22 hand-reared female song sparrows were added to the analysis. General linear mixed models controlling for effects of neophobia and nest of origin indicated that all three song measures were associated with better performance on color reversal and spatial learning but were associated with worse performance on novel foraging and detour-reaching. Overall, the results do not support the hypothesis that learned aspects of song signal cognitive ability.

Social costs are one mechanism whereby reliability in signalling systems can be maintained. We measured the strength of aggressive response to territorial playback to ask whether the reliability of ‘soft song’, a strongly aggressive signal in the song sparrow, Melospiza melodia, is enforced by a social cost in the form of the receiver's aggressive response. We also asked whether this cost is imposed by all receivers, or whether a differential response is found primarily or exclusively among the most aggressive subjects. We first measured the strength of each male's aggressive response to playback on his territory, and then tested his responses to ‘warbled’ soft song and to broadcast song playbacks. While we found substantial variation in individual aggressiveness, nearly all males responded more strongly to warbled soft songs. Thus we provide evidence that warbled soft song imposes a social cost in the form of receiver retaliation, and provide a possible explanation for how the signal's reliability is maintained. Questions about soft song remain, notably why selection should favour low amplitude in vocal signals of aggression in songbirds.

Many organisms use antennae to gather tactile or chemical information from the environment. Crustaceans have two sets of antennae: short antennules (first antennae) and antennae posterior or lateral to them (second antennae) which are often much longer. While the function of crustacean antennae has received some attention, particularly the perception of dispersed chemical signals by the antennules, the role of direct antennal contact during social behavior is less well understood. We compared antennal contact in competitive and pairing interactions in two species of snapping shrimp: Alpheus angulosus and Alpheus heterochaelis. Conspecific interactions involve more antennal contact than heterospecific interactions. Alpheus heterochaelis perform more antennal contact than A. angulosus. Alpheus angulosus females perform more antennal contact in pairing than in competitions; however, A. heterochaelis females and males of both species use antennal contact equally in pairing and competitive interactions. Althou...

In species in which males defend territories for breeding, males may differ in territorial behavior; alternative behaviors among territorial males are not well understood. In our long-term study of partially-migratory song sparrows, we have observed that most territorial males establish territories before females begin nesting and remain site-faithful both within and between breeding seasons; however, some males establish territories later in the season (late establishers) and/or change territory locations either within or between seasons (movers). Whether late establishment or moving are equally successful strategies for territory defense, or best-of-bad-job options, is not known. Here, we compare the frequencies of these behaviors to demographic variables over a 9-yr period and compare lifetime tenure and early season nesting success for males who differ in site fidelity and timing of territory establishment. Across years, late establishing was negatively correlated with the return rate of previously territorial males; moving was positively correlated with the number of occupied territories at the start of the breeding season (territory density). While moving was independent of number of years on territory, late territory establishment only occurred in a male’s first year as a territory holder. Of 88 males, 25% established their first territory late, primarily in undefended space; 31% moved. Late and early establishers did not differ in lifetime tenure; movers, however, had longer lifetime tenure than site-faithful males. Among early establishers, movers and non-movers did not differ in the number of successful early nests/year or number of young fledged/year; among late establishers, however, movers had significantly higher early nesting success by both measures. Late establishers who moved had higher early season nesting success and higher early season nesting success/year than site-faithful early establishers. Thus, individual variation in the timing of territory establishment and site fidelity may be facultative alternative territorial strategies.

Animals in urban habitats face a number of unique stresses, including the necessity of dealing with high levels of human activity. Growing research suggests that: (1) inherent traits, as opposed to learned behavior, influence which species invade urban habitats, and (2) individuals exhibit behavioral syndromes that limit behavioral flexibility. As a result, perhaps only animals with inherently bold personalities successfully settle in areas of high human activity, and such animals may also exhibit correlated variation in other behavioral traits, such as territorial aggression. In this study, we examine boldness and aggression in several urban and rural populations of song sparrows (Melospiza melodia). We found that urban birds were both bolder toward humans and also showed higher levels of aggression. We found a correlation between boldness and aggression in all populations combined, but no correlation within urban populations. Our results agree with other recent studies of song sparrow behavior, suggesting that greater boldness and aggression are general features of urban song sparrow populations, and a lack of a correlation between boldness and aggression in urban habitats is a general phenomenon as well. Urban habitats may select for bold and aggressive birds, and yet the traits can vary independently. These results add to a small number of studies which find that behavioral syndromes break down in potentially high quality habitats.

The role of learning in the development of bird vocalizations other than territorial song is not well studied. The well-known role of direct imitation in the development of territorial song potentially masks the effects of other processes in the development of vocal behaviour. The ‘chick-a-dee’ call of black-capped chickadees is a good system in which to investigate more subtle developmental processes because this call is composed of a small number of distinctive note types. These note types may be classified objectively based on a simple set of acoustic variables, allowing for a quantitative assessment of vocal learning. We raised four groups of black-capped chickadees under different degrees of social and acoustic isolation. We then used a multivariate analysis of the acoustic structure of the introductory call notes (‘A-’ ‘B-’ and ‘C-notes’) to determine how similar the notes produced by these hand-reared birds were to the notes of wild birds. Hand-reared chickadees with greater exposure to normal phonology produced notes of all three note types that were more similar to those of wild birds. Regardless of experience, however, all birds produced A-notes that fell within the normal range of those produced by wild birds. By contrast, the development of normal B- and C-notes appears to be more dependent upon experience. These data suggest that learning may play a different role in the development of different phonological units within one vocalization. Our results also illustrate the importance of considering processes other than simple imitation in the development of avian vocalizations.

Definitions of literacy that focus solely on reading words provide too narrow a framework for many students with severe disabilities. Obtaining information from the environment may be accomplished in a variety of modes, such as visual literacy, which is the ability to discern meaning conveyed through images. A component of visual literacy is picture reading. This study demonstrates the teaching of logo reading through time delay as an additional component of visual literacy. Visual literacy is discussed as a primary means of obtaining information for some students and as an additional means of literacy for students who can also learn to read Words.

Song repertoires are thought to have evolved by sexual selection, with larger repertoires being advantageous in both female choice and territory defence. While most hypotheses of repertoire evolution treat different song types as functionally equal, an alternative hypothesis is that song repertoires evolved to allow song sharing with multiple neighbours. In support of this hypothesis, song sparrows, Melospiza melodia, share high proportions of their repertoires with territorial neighbours in at least three west coast populations in North America, and song sharing is correlated with a territorial advantage in at least two of these. We studied song sharing and territory tenure in an east coast population of song sparrows in North America in which song sharing is significantly less common. We found no evidence for a territorial advantage of whole song sharing in our population. We also found no evidence for a territorial advantage for partial song sharing, even though partial song sharing is as common in our population as whole song sharing is in the west coast populations. Population demographics (such as annual survival and territory density) do not seem sufficient to explain different levels of sharing between populations. Thus, we found no evidence in our population to support song sharing as a target of selection in the evolution of song repertoires.

Access to public community job-training sites can be restricted if students with moderate mental retardation exhibit behavior unacceptable to nondisabled coworkers or the general public. The authors conducted a functional behavioral assessment (FBA) in public community settings to determine the function of each student's problem behavior. They used a multiple-baseline-across-time sample design with behavioral probes embedded within a withdrawal to determine a functional relationship between the independent and dependent variables. The use of self-operated auditory prompts matched to maintaining functions was the function-based intervention. Problem behavior was reduced to criterion for all four students during job training in public community settings.

In a territorial system where males vary in fighting ability or attractiveness to females, not all neighbours should be equally threatening to a territory owner. Selection should act on territory owners to recognize consistent individual differences between neighbours, assess which neighbours represent greater threats, and respond appropriately when challenged. In this study, we examined whether male song sparrows, Melospiza melodia, assess the level of threat posed by individual neighbours and respond differently to neighbours that differ in their level of aggressiveness. Aggressive males should represent a greater threat because they are more likely to engage in fights over territorial boundaries, and may also be preferred by females as extrapair mates. We used playback experiments to assay male territorial aggression, and then to test whether males responded differently to their aggressive and nonaggressive neighbours. We found that male song sparrows respond more strongly to their aggressive neighbours than to their nonaggressive neighbours. Thus, male song sparrows differentiate between neighbours that differ in aggressive response and appear to respond adaptively towards the individual that poses the greatest threat.

In many species, the ability to defend a territory is essential for a male to obtain any reproductive success at all, and even among territorial individuals, variation in the strength of territory defense could have a significant impact on how much reproductive success is obtained. Previous studies have documented consistent individual differences in the vigor with which male song sparrows (Melospiza melodia) defend their territories, as measured by the strength of their reactions to territorial intrusions simulated through song playback. Variation in the strength of defense could reflect intrinsic differences among individuals in their resource holding potential (RHP), or variation in extrinsic factors. In this study, we examined whether variation in the strength of territory defense corresponds to differences in intrinsic factors such as the age or experience of the territory owner, the extrinsic factor of the level of aggression shown by neighbours, or both. Results indicate that males that previously held territories on the study site, regardless of whether they were holding the same territory as the previous year, show higher levels of territory defense than males that are new to the study site, and, assuming that returning males are older males, suggest that age is more important than experience on a specific territory in determining strength of territory defense. In addition, we found evidence that males with high levels of territorial aggression tend to be spatially clustered. The pattern observed suggests that a combination of intrinsic and extrinsic factors contribute to the expression of individual differences in territorial aggression.

This study replicated previous findings that established a five-level, least-to-most prompting system in conjunction with total task presentation as a successful method for teaching students with moderate cognitive disabilities to gain assistance when lost in the community. The present investigation extended previous findings for those students who were unable to identify when they were lost or were unable to independently dial a cell phone to call for assistance. A multiple probe design across students was used to illustrate the effects of intervention on acquisition of cell phone use. The results indicated that all students successfully learned to either answer a ringing cell phone and provide the caller with detailed information about their physical location or use the speed dial function of a cell phone to call for assistance when lost in school and community settings. Limitations and suggestions for future investigations are provided.

Lack (1946) suggested that male songbirds exhibit consistent individual differences in the vigor or manner in which they defend their territories against intrusion. The causes and consequences of such individual variation have not been incorporated into models of territoriality, however, because of a lack of experimental data confirming Lack's suggestion. In this paper, we test the possibility that male song sparrows Melospiza melodia who are successful territory holders differ consistently in the vigor with which they defend their territory, by conducting repeated song playback trials with the same set of territory-holding subjects across a breeding season. We found only relatively weak seasonal trends in responsiveness: the amount birds sang in response to playback increased significantly across the breeding season and responsiveness was generally lower when a male's social mate was egg laying. By contrast, we found extensive variation among males in how closely they approached a simulated territorial intrusion. These individual differences remained significantly consistent across four rounds of playback trials that spanned the breeding season as determined by Kendall's coefficient of concordance. Our results confirm that some individual song sparrows are consistently more vigorous than others in territory defense, at least in one conspicuous aspect of their behavior, and suggest that further work is needed to understand the nature and consequences of variation in patterns of defense among successful territory holders.

This study examined the acquisition of cell phone use by middle school students with moderate cognitive disabilities. While in school and community settings, students were taught to identify when they were lost and follow a set of procedures for using a cell phone to call for assistance. Through the use of a five-level prompting system in conjunction with total task presentation, students acquired these new safety skills. A series of concurrent multiple-probe designs across groups was used to evaluate cell phone use and to permit generalization from school to community settings. For four participants, generalization was also made to a second phone call recipient. Results indicated that all students successfully acquired the skills of identifying when they were lost and then using a cell phone to call for assistance.

Song learning in oscine birds is often defined solely as a process of song imitation; nonetheless, not all songs produced by laboratory-tutored birds are imitations of the model songs. If song learning were strictly a process of imitation, these non-imitated songs (inventions) would be expected to contain no learned attributes. To determine whether species-typical song attributes can be learned in the absence of imitation, we compared the imitations and inventions of laboratory-tutored nightingales (Luscinia megarhynchos B.) with the songs of wild nightingales and the songs of laboratory-reared, untutored nightingales. The species-typical song attribute measured was stereotypy. We quantified stereotypy by four variables: (1) percentage of notes shared between two renditions of the same song type (2) difference in repetition rates of the same trill in two renditions of the same song type (3) acoustic similarity of the same note in two renditions of the same song type, and (4) acoustic similarity of the same note repeated within a trill. Wild songs and imitated songs were significantly more stereotypical than the songs of untutored birds for all measures. For the percentage of notes shared (1), and the acoustic similarity of notes in two renditions of the same song type (3), invented songs did not differ from the songs of untutored birds, suggesting that imitation is necessary for the acquisition of these song characteristics. However, invented songs were significantly more stereotypical than the songs of untutored birds for measures of stereotypy related to trills (2 and 4), and neither imitated nor invented songs differed significantly from the songs of wild birds in terms of trill rate stereotypy (2), Thus, it appears that the process of learning to produce trills may differ from the process of learning non-repetitive song components: increased stereotypy in trills occurs even when the trills themselves are not copied from song models. Strict imitation does not fully account for the acquisition of some learned song attributes.

Background: Focal adhesion Kinase (FAK) is a nonreceptor protein tyrosine kinase that is overexpressed in tumors and plays a significant role in tumor survival and metastasis. The purpose of the study is to perform correlation of FAK expression with patient prognostic factors using tissue microarrays (TMA) samples. Methods: We analyzed FAK expression by immunohistochemical staining in 196 breast primary tumor samples from stage II-IV patients and in 117 metastatic tissues matched to the primary tumors using TMA that were stained with FAK monoclonal antibody. Results: High FAK expression in primary tumors was associated with a younger age of patients (p = 0.033), lymphovascular invasion (p = 0.001) and with the triple-negative phenotype (p = 0.033). FAK expression in 117 metastatic tissues positively correlated with FAK expression in matched primary tumors by Spearman correlation analysis. In addition, a strong positive correlation was observed between high FAK expression and shorter overall survival and progression free survival in patients with metastatic tumors. Conclusions: The data demonstrate a high potential for FAK as a therapeutic target, especially in triple-negative breast cancer patients with high FAK expression.