Quantifying effectiveness and best practices for bumblebee identification from photographs.

Understanding pollinator networks requires species level data on pollinators. New photographic approaches to identification provide avenues to data collection that reduce impacts on declining bumblebee species, but limited research has addressed their accuracy. Using blind identification of 1418 photographed bees, of which 561 had paired specimens, we assessed identification and agreement across 20 bumblebee species netted in Montana, North Dakota, and South Dakota by people with minimal training. An expert identified 92.4% of bees from photographs, whereas 98.2% of bees were identified from specimens. Photograph identifiability decreased for bees that were wet or matted; bees without clear pictures of the abdomen, side of thorax, or top of thorax; bees photographed with a tablet, and for species with more color morphs. Across paired specimens, the identification matched for 95.1% of bees. When combined with a second opinion of specimens without matching identifications, data suggested a similar misidentification rate (2.7% for photographs and 2.5% specimens). We suggest approaches to maximize accuracy, including development of rulesets for collection of a subset of specimens based on difficulty of identification and to address cryptic variation, and focused training on identification that highlights detection of species of concern and species frequently confused in a study area.

Neoadjuvant weekly paclitaxel and carboplatin with trastuzumab and pertuzumab in HER2-positive breast cancer: a Brown University Oncology Research Group (BrUOG) study.

PURPOSE: In HER2-positive breast cancer (HER2+ BC), neoadjuvant chemotherapy (NACT) with dual HER2-targeted therapy achieves high pathologic complete response (pCR) rates. Anthracycline-free NACT regimens avoid toxicities associated with anthracyclines, but every 3-week TCHP also has substantial side effects. We hypothesized that a weekly regimen might have equivalent efficacy with less toxicity; we also investigated whether poorly responding patients would benefit from switching to AC. METHODS: Patients with clinical stage II-III HER2+ BC received weekly paclitaxel 80 mg/m2 and carboplatin AUC2 with every 3-week trastuzumab and pertuzumab (wPCbTP), with the option of splitting the pertuzumab loading dose. After 12 weeks, responding patients continued wPCbTP for another 6 weeks, while non-responders switched to AC. Dose modifications and post-op therapy were at investigator discretion. RESULTS: In 30 evaluable patients, the pCR rate was 77% (95% CI 58-90%); 12/14 (86%) in ER-negative and 11/16 (69%) in ER-positive. Only two patients transitioned to AC for non-response, of which one achieved pCR. There were no episodes of febrile neutropenia or grade ≥ 3 peripheral neuropathy, though several patients who continued wPCbTP stopped before week 18. Split-dose pertuzumab was associated with less grade ≥ 2 diarrhea (40%) than the standard loading dose (60%). CONCLUSION: pCR rates with our regimen were as high as reported with TCHP with fewer grade ≥ 3 toxicities, though diarrhea remains a concern. Too few patients had a suboptimal response to adequately test switching to AC. The wPCbTP regimen should be considered an alternative to TCHP as neoadjuvant therapy for HER2+ BC. TRAIL REGISTRATION: ClinicalTrials.gov identifier: NCT02789657.

A comparative analysis of lobular and ductal carcinoma of the breast: presentation, treatment, and outcomes.

BACKGROUND: Compared with invasive ductal carcinoma, invasive lobular carcinoma of the breast is considered by many to be a more indistinct and multicentric form of cancer that is detected later and is treated less optimally by breast-preservation techniques. This study analyzed the presentation, treatment trends, and survival rates of women who had invasive lobular and ductal breast carcinoma. The objective was to determine the utility of breast-preservation therapy for invasive lobular carcinoma by analysis of historic data on tumor features and survival. STUDY DESIGN: Data on 291,273 women diagnosed with invasive carcinoma between 1985 and 1993 were obtained from the National Cancer Data Base. Analysis included the patient's age at diagnosis; tumor histology, anatomic site, diameter, grade, and stage; treatment; and disease status 5 years after diagnosis. RESULTS: The mean patient age at diagnosis was 61.0 years for invasive ductal carcinoma, 63.0 years for invasive lobular carcinoma, and 60.6 years for tumors with combined histology. The anatomic location, tumor diameter, and tumor grade were similar for each histotype. Breast-preservation therapy was less frequent for invasive lobular carcinoma. The 5-year overall survival and local disease-free survival rates for women treated with breast preservation were similar for invasive ductal carcinoma (84% overall survival; 97% disease-free survival) and invasive lobular carcinoma (87% overall survival; 98% disease-free survival). CONCLUSIONS: Invasive lobular carcinoma presents with a similar age distribution, anatomic subsite, diameter, and grade as invasive ductal carcinoma. Breast preservation is selected less commonly for women who have invasive lobular carcinoma, but this choice of therapy does not compromise the disease-free or overall survival status of this group of patients.

Surgery for early and minimally invasive breast cancer.

An increasing number of early and minimally invasive breast cancers are being detected by mammography screening. The role of image-guided core biopsy in nonpalpable breast lesion evaluation is being defined, with issues of cost-effectiveness, diagnostic accuracy, and ability to direct definitive management remaining to be elucidated. The long-term efficacy of breast conservation therapy for early breast cancer continues to be confirmed. Review of special issues in breast conservation therapy demonstrates no benefit to eliminating adjunctive radiation therapy or to limiting therapy in the elderly patient with breast cancer. The distribution of cancer in the duct-lobular system indicates the potential for radical segmental extension. It also influences local recurrence following breast-conserving surgery. Sentinel lymph node dissection may supplant standard axillary dissection as a major prognostic determinant of metastatic disease. These areas of scientific investigation continue to define the role of surgery in early and minimally invasive breast cancer.

Comorbidity risk parameters associated with advanced breast cancer and systemic disease. Management of nonbreast disease.

The comorbidity risks associated with advanced breast cancer and systemic disease have been outlined under three broad categories: the local or regional consequences of advanced local breast cancer and its management, systemic sequelae of adjuvant chemotherapy and radiotherapy, and systemic sequelae of metastatic breast disease. Awareness of the potential risks, early detection, and active intervention may diminish the adverse consequences of comorbidities in breast cancer and improve the patient's quality of life.

The renal effects of low-dose dopamine in thermally injured patients.

The renal effects of low-dose dopamine (LDD) therapy in hyperdynamic thermally injured patients are unknown. We investigated the renal effects of LDD in ten burn patients (mean +/- SEM age and % total body surface burned: 30.2 +/- 3.3 years and 53.4% +/- 7%) and six controls (mean age; 20.2 +/- 0.5 years). Administration of LDD significantly increased glomerular filtration rate, effective renal plasma flow, sodium excretion, and urine flow in the controls and effective renal plasma flow, urine flow, heart rate, and cardiac index in the patients. The chronotropic effect of dopamine appears to be a principal contributor to the patients' increased effective renal plasma flow. Sodium excretion was increased by LDD only in the patients in whom the predopamine sodium excretion exceeded 5 mEq/h. Lack of a consistent natriuretic effect and the consistent chronotropic effect suggest that the routine use of low-dose dopamine in burn patients is unwarranted. The side effects that attend the desired response determine clinical use, i.e., the potential for blood flow redistribution and increased cardiac work demands must be balanced against increased renal plasma flow and natriuresis.

Prophylactic use of high-frequency percussive ventilation in patients with inhalation injury.

Death and the incidence of pneumonia are significantly increased in burn patients with inhalation injury, despite application of conventional ventilatory support techniques. The effect of high-frequency percussive ventilation on mortality rate, incidence of pulmonary infection, and barotrauma were studied in 54 burn patients with documented inhalation injury admitted between March 1987 and September 1990 as compared to an historic cohort treated between 1980 and 1984. All patients satisfied clinical criteria for mechanical ventilation. High-frequency percussive ventilation was initiated within 24 hours of intubation. The patients' mean age and burn size were 32.2 years and 47.8%, respectively (ranges, 15 to 88 years; 0% to 90%). The mean number of ventilator days was 15.3 +/- 16.7 (range, 1 to 150 days), with 26% of patients ventilated for more than 2 weeks. Fourteen patients (25.9%) developed pneumonia compared to an historic frequency of 45.8% (p less than 0.005). Mortality rate was 18.5% (10 patients) with an expected historic number of deaths of 23 (95% confidence limits of 17 to 28 deaths). The documented improvement in survival rate and decrease in the incidence of pneumonia in patients treated with prophylactic high-frequency ventilation (HFV), as compared to a cohort of patients treated in the 7 years before the trial, indicates the importance of small airway patency in the pathogenesis of inhalation injury sequelae and supports further use and evaluation of HFV.

Relationship of transfusion and infection in a burn population.

The relationship between the number of red blood cell transfusions and major infectious complications was evaluated in 594 thermal injury patients admitted between 1982 and 1986 who had burns over 10% or more of total body surface area and survived more than 10 days. The mean age of this group was 32.9 years, with a mean burn size of 36% of total body surface area; 83% were male. Of the 594 patients, 23.7% died and 38.7% had documented inhalation injury. The mean number of red blood cell transfusions received was 19.7, with a range of 0 to 201. Two hundred fourteen patients (36%) had major infectious complications, defined as pneumonia or invasive burn wound infection. A cross-tabulation of predicted mortality, number of transfusions, and infectious complications revealed a significant positive correlation between transfusion number and infectious complications in patients with predicted mortalities between 10 and 70%. Per cent total burn, patient age, presence of inhalation injury, and number of transfusions were identified by discriminant function analysis as significant variables (p less than 0.05) in discriminating between patients with and without infections (85% accuracy). Logistic regression analysis confirmed the above findings, showing a relationship between the number of transfusions received and infectious morbidity which was independent of age or burn size, but no significant relationship between number of transfusions and mortality.

High-frequency percussive ventilation in patients with inhalation injury.

Inhalation injury complicated by bacterial pneumonia is now one of the primary causes of morbidity and mortality in patients with thermal injury. We have investigated the use of high-frequency percussive ventilation (HFPV) as a means of ventilatory support for these patients. We propose that high-frequency ventilation may decrease the incidence of pulmonary infection following inhalation injury and decrease the incidence of iatrogenic barotrauma caused by conventional ventilation. High-frequency ventilation was instituted initially as salvage therapy in a group of five patients. In each case, normocapnia or arterial pO2 saturation of greater than 90% on a FIO2 of 60% or less was achieved with high-frequency ventilation but not with conventional ventilation. A second group of ten patients was prospectively entered into a study on the use of HFPV in patients with inhalation injury. One patient was removed from the study, and one patient was unable to be ventilated because of severely noncompliant lungs. Eight patients with a mean age of 29 years and a mean burn size of 38% of the total body surface completed the protocol. All patients survived, two developed pneumonia, and one developed subcutaneous emphysema. These results suggest that HFPV is effective in the treatment of patients with severe inhalation injury.

Fluid resuscitation of infants and children with massive thermal injury.

Age-related limitations of physiologic reserve in burned children make adequacy of intravenous fluid resuscitation critical. To quantify fluid requirements, the medical records of all children admitted to the Army Institute of Surgical Research from 1980 to 1986 whose weight was 25 kilograms or less and burn size was 25% or greater were reviewed to quantify fluid requirements. Forty-three children ranging in age from 1.5-108 months (means 26 +/- 22 m), with 25-89% total body surface burned (TBSB) (means 41.7% +/- 14.6%), met inclusion criteria. The average total volume of fluid received during the first 24 hours was 6.3 +/- 2.2 cc/kg/% TBSB. The net volume of resuscitation fluid, total volume less the calculated maintenance fluid requirements, was 3.91 +/- 2.2 cc/kg/% TBSB. Univariate analysis and linear regression of independent variables including age, weight, per cent full thickness, and inhalation injury revealed each had no significant influence on the volume of resuscitation. We recommend supplying maintenance volume and initiating burn resuscitation at 3 cc/kg/% TBSB.

Regulation of microvillus structure: calcium-dependent solation and cross-linking of actin filaments in the microvilli of intestinal epithelial cells.

The bundle of filaments within microvilli of intestinal epithelial cells contains five major proteins including actin, calmodulin, and subunits of 105-, 95-, and 70-kdaltons. It has been previously shown (Howe, C. L., M. S. Mooseker, and T. A. Graves. 1980. Brush-border calmodulin: a major component of the isolated microvillus core. J. Cell Biol. 85: 916-923) that the addition of Ca++ (> 10(-6) M) to microvillus cores causes a rapid, drastic, but at least partially reversible disruption of this actin filament bundle. High-speed centrifugation of microvillus cores treated with Ca++ indicates that several core proteins are solubilized, including 30-50% of the actin and calmodulin, along with much of the 95- and 70-kdalton subunits. Gel filtration of such Ca++ extracts in the presence and absence of Ca++ indicates that microvillar actin "solated" by Ca++ is in an oligomeric state probably complexed with the 95-kdalton subunit. Removal of Ca++ results in the reassembly of F-actin, probably still complexed with 95-kdalton subunit, as determined by gel filtration, cosedimentation, viscometry, and electron microscopy. The 95-kdalton subunit (95K) was purified from Ca++ extracts by DEAE-Sephadex chromatography and its interaction with actin characterized by viscometry, cosedimentation, and EM in the presence and absence of Ca++. In the presence, but not absence, of Ca++, 95K inhibits actin assembly (50% inhibition at 1:50-60 95K to actin) and also reduces the viscosity of F-actin solutions. Similarly, sedimentation of actin is inhibited by 95K, but a small, presumably oligomeric actin- 95K complex formed in the presence of Ca++ is pelletable after long-term centrifugation. In the absence of Ca++, 95K cosediments with F-actin. EM of 95K-actin mixtures reveals that 95K "breaks" actin into small, filamentous fragments in the presence of Ca++. Reassembly of filaments occurs once Ca++ is removed. In the absence of Ca++, 95K has no effect on filament structure and, at relatively high ratios (1:2-6) of 95K to actin, this core protein will aggregate actin filaments into bundles.

Brush-border calmodulin. A major component of the isolated microvillus core.

Calmodulin is present in brush borders isolated from intestinal epithelial cells and is one of the major components of the microvillar filament bundle. Calmodulin was purified from either demembranated brush borders or microvilli by a simple boiling procedure. The boiled supernate derived from the microvillus cores contained one major polypeptide of 20,000 daltons. The supernate from the brush-border preparation contained the 20,000-dalton subunit and a second protein of 30,000 daltons. The 20,000-dalton subunit has been identified as calmodulin by several criteria: (a) heat resistance, (b) comigration with brain calmodulin on alkaline urea gels and SDS gels, both cases in which the 20,000-dalton protein, like calmodulin, exhibits a shift in electrophoretic mobility in the presence of Ca++, and (c) 4--5-fold activation of 3',5'-cyclic nucleotide phosphodiesterase in the presence but not the absence of Ca++. With a cosedimentation assay it was determined that brush-border calmodulin does not bind directly to actin. In the presence of Ca++ (greater than 5 x 10(-7) M) there was a partial release of calmodulin from the microvillus core, along with a substantial conversion of microvillus actin into a nonpelletable from. The dissociation of calmodulin was reversed by removal of Ca++. If microvillus cores were pretreated with phalloidin, the Ca++-induced solubilization of actin was prevented, but the partial dissociation of calmodulin still occurred. The molar ratio of calmodulin:actin is 1:10 in the demembranated brush border and 1:2-3 in the microvillus core. No calmodulin was detected in the detergent-solubilized brush-border membrane fraction.