Normal Growth Rate Research Articles

Characterization of resistance to a potent d-peptide HIV entry inhibitor

BackgroundPIE12-trimer is a highly potent d-peptide HIV-1 entry inhibitor that broadly targets group M isolates. It specifically binds the three identical conserved hydrophobic pockets at the base of the gp41 N-trimer with sub-femtomolar affinity. This extremely high affinity for the transiently exposed gp41 trimer provides a reserve of binding energy (resistance capacitor) to prevent the viral resistance pathway of stepwise accumulation of modest affinity-disrupting mutations. Such modest mutations would not affect PIE12-trimer potency and therefore not confer a selective advantage. Viral passaging in the presence of escalating PIE12-trimer concentrations ultimately selected for PIE12-trimer resistant populations, but required an extremely extended timeframe (> 1 year) in comparison to other entry inhibitors. Eventually, HIV developed resistance to PIE12-trimer by mutating Q577 in the gp41 pocket.ResultsUsing deep sequence analysis, we identified three mutations at Q577 (R, N and K) in our two PIE12-trimer resistant pools. Each point mutant is capable of conferring the majority of PIE12-trimer resistance seen in the polyclonal pools. Surface plasmon resonance studies demonstrated substantial affinity loss between PIE12-trimer and the Q577R-mutated gp41 pocket. A high-resolution X-ray crystal structure of PIE12 bound to the Q577R pocket revealed the loss of two hydrogen bonds, the repositioning of neighboring residues, and a small decrease in buried surface area. The Q577 mutations in an NL4-3 backbone decreased viral growth rates. Fitness was ultimately rescued in resistant viral pools by a suite of compensatory mutations in gp120 and gp41, of which we identified seven candidates from our sequencing data.ConclusionsThese data show that PIE12-trimer exhibits a high barrier to resistance, as extended passaging was required to develop resistant virus with normal growth rates. The primary resistance mutation, Q577R/N/K, found in the conserved gp41 pocket, substantially decreases inhibitor affinity but also damages viral fitness, and candidate compensatory mutations in gp160 have been identified.

Endogenous growth, capital accumulation and Thirlwall's dynamics: the case of Latin America

The article draws together the analyses of the interaction between economic capacity (ec), the endogeneity of the natural growth rate (gn) and the growth rate consistent with balance-of-payments equilibrium (gtb) that constrains economic activity. We identify two possible scenarios: the self-correcting scenario where gtb is more elastic than the normal natural rate of growth (gnn) vis-à-vis ec, and the self-aggravating scenario where gnn is more elastic than gtb with respect to ec. We empirically assess our central tenet (ec is a determinant of the relations between gtb, gw and gnn) for the cases of Argentina, Brazil, Chile and Mexico, and found that, in all countries, the relationships between ec and gnn and between ec and gtb are positive, except in the case of Argentina where the relation between gtb and ec was negative in the sub-period 1975–1990.

Impact of ferumoxytol magnetic resonance imaging on the rhesus macaque maternal-fetal interface†.

Ferumoxytol is a superparamagnetic iron oxide nanoparticle used off-label as an intravascular magnetic resonance imaging (MRI) contrast agent. Additionally, ferumoxytol-uptake by macrophages facilitates detection of inflammatory sites by MRI through ferumoxytol-induced image contrast changes. Therefore, ferumoxytol-enhanced MRI holds great potential for assessing vascular function and inflammatory response, critical to determine placental health in pregnancy. This study sought to assess the fetoplacental unit and selected maternal tissues, pregnancy outcomes, and fetal well-being after ferumoxytol administration. In initial developmental studies, seven pregnant rhesus macaques were imaged with or without ferumoxytol administration. Pregnancies went to term with vaginal delivery and infants showed normal growth rates compared to control animals born the same year that did not undergo MRI. To determine the impact of ferumoxytol on the maternal-fetal interface (MFI), fetal well-being, and pregnancy outcome, four pregnant rhesus macaques at ~100 gestational day underwent MRI before and after ferumoxytol administration. Collection of the fetoplacental unit and selected maternal tissues was performed 2-3days following ferumoxytol administration. A control group that did not receive ferumoxytol or MRI was used for comparison. Iron levels in fetal and MFI tissues did not differ between groups, and there was no significant difference in tissue histopathology with or without exposure to ferumoxytol, and no effect on placental hormone secretion. Together, these results suggest that the use of ferumoxytol and MRI in pregnant rhesus macaques does not negatively impact the MFI and can be a valuable experimental tool in research with this important animal model.

Three-Dimensional Experimental Study of Compressibility Effects on Turbulent Free Shear Layers

Turbulent planar mixing layers of varying compressibility levels have been studied experimentally via flow visualization techniques, pressure measurements, and stereoscopic particle image velocimetry. The experiments were conducted for five different convective Mach numbers (, 0.38, 0.55, 0.69, and 0.88). For each case, inflow conditions were thoroughly documented with planar particle image velocimetry. Results indicate that the incoming boundary layers are thin (less than or equal to 9% of channel height) and fully developed. Schlieren and Mie scattering images show that for increased levels of compressibility the large-scale turbulent structures are less organized, and the Brown–Roshko rollers that are dominant in the incompressible case are not found. Three-component velocity measurements made on the spanwise-central plane () via stereo particle image velocimetry confirm the reduction in the normalized growth rate with increasing that has been agreed upon widely. Turbulence statistics results show that the peak (normalized) streamwise normal kinematic Reynolds stress remains constant, while the spanwise normal, transverse normal, and primary shear kinematic Reynolds stress peak values all decrease with increasing . In addition, knowledge of all three normal stress components allows for the calculation of the full Reynolds stress anisotropy tensor, a novel result for compressible mixing layer experiments. Each anisotropy component remains constant across the shear layer center. As compressibility in the mixing layer increases, the streamwise normal stress anisotropy increases, transverse and spanwise normal stress anisotropies decrease, and the shear stress anisotropy remains constant. Conversely, the Reynolds stress correlation coefficient remains constant across all cases at approximately 0.47.

P81. Mitigating distal migration of apex by reducing apical vertebral wedging alongside guided growth technique: active apex compression technique

BACKGROUND CONTEXT Distraction system either cause immense morbidity to the patient by several consecutive and invasive distraction episodes or are extremely expensive and not immediately affordable for a major of patients worldwide. SHILLA procedure, although it doesn't result in consecutive surgeries, there still remains severe complications of loss of correction or reappearance of deformity through crankshafting or adding-on (for eg, distal migration). The main culprit of this complication is the vertebral growth anteriorly at the apex, which mostly remains unmodulated by static fusion posteriorly. The current study presents a modified approach to SHILLA that could help dynamically remodulate, ie, reverse modulate, the apex of the deformity. PURPOSE To determine if active remodulation in the apex of the curve is possible in scoliosis and kyphoscoliosis patients, using a modified SHILLA; APC technique for guided growth. STUDY DESIGN/SETTING A retrospective cohort with paired control group. PATIENT SAMPLE Twenty patients paired as follows: (1) 20 adjacent vertebra; (2) 20 compressed wedged vertebra. OUTCOME MEASURES The convex and concave heights of the wedged vertebra were recorded at the time of the surgery and at follow up, both using CT. For comparison a control vertebra was chosen, whose physical dimensions were in par with the wedged vertebra to record the normal growth rate of the particular duration of follow up and age. METHODS The study consisted of 20 patients with either scoliosis or kyphoscoliosis undergoing an index surgery or revision surgery and demonstrating a clear radiographic evidence of vertebral wedging at the apex. All patients were under 8 years of age with Risser less than or equal to 2, and the major Cobb's angle more than 40°. The surgical procedure involved was a modified version of SHILLA, either using rod to screw (SHILLA screws from Medtronic) sliding mechanism or the analogous rod to domino (4.5 mm rod in 5.5 mm domino) sliding mechanism. In this modified technique, the most wedged vertebra was selected followed by insertion of pedicle screws in the convex side of the vertebrae above and below the wedged one. No screws were put on the concave side of the apex. All surgeries were performed under intraoperative neuromonitor and C-arm. Additionally, no cast or brace were used for these patients postoperatively. The patients were followed for a period of at least 8 months. The convex and concave heights of the wedged and control vertebrae were recorded at the time of the surgery and at follow-up, both using CT. The main difference between a wedged vertebra and the control vertebra was the compression forces being applied at the convex end for the wedged vertebra, with the expectations of reducing local longitudinal growth rate. In the control group no such forces were applied. RESULTS The wedged vertebra demonstrated a 17% average increase (p=0.00014) in the proportion of concave to convex heights ratio, whereas the control vertebra didn't show any relative change in the wedged vertebra heights at the follow ups. CONCLUSIONS Active apex correction, instead of apical fusion in SHILLA remodulates the apex vertebra, which may in turn help mitigate loss of correction on long term due to crankshafting and adding-on. FDA DEVICE/DRUG STATUS Unavailable from authors at time of publication.

Donkey\u2019s Milk in the Management of Children with Cow\u2019s Milk protein allergy: nutritional and hygienic aspects

BackgroundThe therapeutic strategy for children with cow’s milk allergy (CMA) consists in the elimination of cow’s milk (CM) from their diet. Donkey’s milk (DM) has been reported to be an adequate alternative, mainly to his nutritional similarities with human milk (HM) and excellent palatability. The aim of present prospective study was to evaluate the nutritional impact of DM on the diet of children with CMA in term of children growth.MethodsBefore the nutritional trial on children and during the study the health and hygiene risks and nutritional and nutraceuticals parameters of DM were monitored. Children with CMA were identified by the execution of in vivo and in vitro tests for CM and subsequent assessment of tolerability of DM with oral food challenge (OFC). Finally, we prescribed DM to a selected group of patients for a period of 6 months during which we monitored the growth of children. A total of 81 children, 70 with IgE mediated cow’s milk protein allergy (IgE-CMPA) and 11 with Food Protein Induced Enterocolitis Syndrome to CM (CM-FPIES), were enrolled.ResultsSeventy-eight out of 81 patients underwent the OFC with DM and only one patient with IgE-CMPA (1.5 %) reacted. Twenty-two out of 81 patients took part of the nutritional trial. All the 22 patients took and tolerated the DM, moreover DM did not change the normal growth rate of infants.ConclusionsIn conclusion, DM resulted safe in term of health and hygiene risks and nutritionally adequate: no negative impact on the normal growth rate of children was assessed. Therefore, it may be a suitable alternative for the management of IgE mediated CMA and FPIES, also in the first 6 months of life, if adequately supplemented.

PENGARUH DEBT TO EQUITY RATIO (DER), UKURAN PERUSAHAAN(SIZE), NET PROFIT MARGIN (NPM) DAN RETURN ON ASSETS (ROA)TERHADAP PERATAAN LABA (IS)

Income smoothing is a phenomenon used by management with the aim of reducing variability in earnings over a certain period or in a period, which leads to the expected level of reported earnings. Efforts to reduce the fluctuation of earnings is a form of profit manipulation so that the amount of profit in a period is not too different from the amount of profit in the previous period. Therefore, income smoothing involves the use of certain techniques to reduce or increase the amount of profit of a period with the amount of profit of the previous period. However, this effort is not to make a profit of a period equal to the amount of the profit of the previous period, because in reducing the fluctuation of profit also considered the normal growth rate expected in that period.

Normal Diameter and Growth Rate of Infrarenal Aorta and Common Iliac Artery in Chinese Population Measured by Contrast-Enhanced Computed Tomography

In previous studies, the growth rate and diameter of infrarenal aorta was paucity of Chinese data. This study aim to evaluate the normal inside diameter and growth rate of the infrarenal aorta and common iliac artery in Chinese adult population and to explore the relationship between related factors and the arterial diameter. A retrospective study was performed among 1,340 hospitalized adult patients undergoing contrast-enhanced abdominopelvic computed tomography from April 2017 to March 2018 in our hospital. The inside diameter of infrarenal aorta and common iliac artery was measured. Linear regression was used to analyze the relationship between various factors and arterial diameter. Kruskal-Wallis H test was used to analyze the relationship between the arterial diameter and age. Mann-Whitney U test was used to analyze the relationship between arterial diameter and sex. The inside diameter of the infrarenal aorta was 16.49±2.12mm in male patients and 14.50±1.73mm in female patients. In male patients, the right common iliac artery was 9.77±1.75mm, and the left was 9.65±1.76mm. In female patients, the right common iliac artery was 8.59±1.31mm and the left was 8.45±1.28mm. Comparing the oldest group with the youngest, the infrarenal aortic diameter increased 27.32% in male patients and 30.11% in female patients. Right common iliac artery increased 25.13% in male patients and 30.30% in female patients. Left common iliac artery increased 25.19% in male patients and 34.26% in female patients. The growth rate increased at the beginning, reached its peak at the age of 50-60years, and then decreased. Multiple linear regression analysis results showed that sex, age, body surface area (BSA), hypertension, and cancer were significantly correlated with the diameter of infrarenal aorta. Sex, age, BSA, and hypertension were correlated with the diameter of right common iliac artery. Sex, age, BSA, and cancer were correlated with the diameter of left common iliac artery. The diameter of the infrarenal abdominal aorta and common iliac artery of Chinese people is smaller than that of other countries. The abdominal aortic aneurysm (AAA) guideline might be developed based on the basic diameter to suit patients with AAA and a different basic abdominal aortic diameter. Artery diameter was increased by the age, and female patients have smaller diameter and larger arterial growth rate than male patients. Female and young patients with AAA may choose more oversize grafts for endovascular AAA repair.

Improving Growth Velocity in Very Low Birth Weight Infants: a Quality Improvement Project

Background: Inadequate nutritional support in very low birth weight (VLBW) infants results in growth failure and poor neurodevelopmental outcomes. Early optimization of nutrition, especially protein intake, is an important strategy to achieve normal postnatal growth. Most infants postnatally demonstrate weight loss, do not sustain normal intrauterine growth rates once growth resumes, and do not achieve normal growth by term gestation. Identified reasons include: delays in achieving adequate nutrition, inadequate protein intake, late initiation of enteral feedings, use of unfortified human milk, and variability in practices between providers. To improve growth …

Abstract 4245: Drug response metrics and pharmacological profiling using the OncoPanel™ cell-based profiling service

Abstract High-throughput, cell-based drug screening platforms are essential to guiding drug discovery programs and can rapidly generate large amounts of pharmacological information about a candidate drug. Drug response data combined with genomic and phenotypic information enables an exploratory analysis that may reveal potential associative biomarkers and deepen the pharmacological profile of a drug. The choice of metric is an important consideration as different metrics will reflect different underlying biological mechanisms. Normalized growth rate inhibition (GR) metrics offer an alternative summary of drug response, and may eliminate bias associated with experimental factors that contribute to variation in growth rate. In these sets of experiments, we used drug response data, generated with the Eurofins OncoPanel cell-based profiling service, to investigate a variety of standard of care compounds against a cell line panel selected for variation in growth rate, cancer indication, cancer hallmark mutational status, and baseline apoptosis rate. Seeding density and incubation times were systematically varied to investigate the relationship between end-point confluence, kinetics, genotype, apoptotic priming, and the robustness of different dose response metrics. Based on these results, select cell lines and standard of care compounds were included in a series of drug-drug combination studies to further investigate the biological underpinning of synergy, at the same time exploring how alternative metrics influence pharmacological profiling experiments. Compounds were added to cells in 10 half-log dilutions, in triplicate, and allowed to incubate for 1, 3, 5, or 10 days. Initial cell density conditions were varied to allow for a final targeted confluence of 30, 80, or 100%. A time-zero plate was fixed at compound addition to allow for GR calculations. Following incubation, the cells were fixed, stained with DAPI, and imaged with a high-content imaging system. Cell proliferation dose response curves were fitted using a 4-parameter log-logistic model with custom curve-fitting software. These data were re-analyzed using the GRmetrics package in R to determine the GR metrics. Summary parameters, including IC50, GR50, and GRmax, were compared in subsequent analyses. For the combination analyses, a 9 x 9 concentration matrix design was used, and Bliss analysis was conducted to assess synergy. Single parameters are typically used to summarize drug responses in cell lines, which are used to classify cell lines as sensitive or resistant to the drug, impacting the interpretation of downstream analyses. We examined the robustness of different parameters, under various growth conditions for select standard of care compounds against a panel of cancer cell lines, and investigated their interpretation on biomarker and drug-drug combination analyses. Citation Format: Charles R. Wageman, Lee R. Cavedine, Vanessa Norman, Natiya Robinson, Tracy Lu, Victoria McBain, Joseph Murphy, Kayla Stehle, Steven M. Garner, Alyssa M. Croff, Brogan A. Epkins, Kristin Dempsey, Alastair J. King, Jesse J. Parry. Drug response metrics and pharmacological profiling using the OncoPanel™ cell-based profiling service [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 4245.

Recreational therapy workforce: An update

This article explores why there is a decline in the growth rate of recreational therapy while there are substantial increases in other therapy professions and recreation workers. The growth numbers of the recreational therapy profession are at a normal growth rate of 7 percent for all occupations according to the Bureau of Labor Statistics (BLS). The growth rate from the Occupational Handbook for 2010-2011 projected a growth rate of 15 percent. The growth rates for occupational therapy, “therapists, all others” and recreation workers are higher than recreational therapy projected by BLS at 20 percent or higher. Possible considerations as to causes for this change in growth are proposed.

Autologous stem cell therapy for hypoplastic left heart syndrome: Safety and feasibility of intraoperative intramyocardial injections

ObjectivesStaged surgical palliation for hypoplastic left heart syndrome results in an increased workload on the right ventricle serving as the systemic ventricle. Concerns for cardiac dysfunction and long-term heart failure have generated interest in first-in-infant, cell-based therapies as an additional surgical treatment modality. MethodsA phase 1 clinical trial was conducted to evaluate the safety and feasibility of direct intramyocardial injection of autologous umbilical cord blood-derived mononuclear cells in 10 infants with hypoplastic left heart syndrome at the time of stage II palliation. ResultsAll 10 patients underwent successful stage II palliation and intramyocardial injection of umbilical cord blood-derived mononuclear cells. Operative mortality was 0%. There was a single adverse event related to cell delivery: An injection site epicardial bleed that required simple oversew. The cohort did not demonstrate any significant safety concerns over 6 months. Additionally, the treatment group did not demonstrate any reduction in cardiac function in the context of the study related intramyocardial injections of autologous cells. ConclusionsThis phase 1 clinical trial showed that delivering autologous umbilical cord blood-derived mononuclear cells directly into the right ventricular myocardium during planned stage II surgical palliation for hypoplastic left heart syndrome was safe and feasible. Secondary findings of preservation of baseline right ventricular function throughout follow-up and normalized growth rates support the design of a phase 2b follow-up trial.

Height Velocity Defined Metabolic Control in Children With Congenital Adrenal Hyperplasia Using Urinary Steroid GC-MS Analysis.

Treatment of children with classic congenital adrenal hyperplasia (CAH) with glucocorticoids is a difficult balance between hypercortisolism and hyperandrogenism. Biochemical monitoring of treatment is not well defined. Achievement of a normal growth rate is the most important therapeutic goal. We retrospectively evaluated 123 24-hour gas chromatography-mass spectrometry urinary steroid metabolome analyses together with their corresponding 1-year height velocity (HV) z scores in 63 prepubertal children aged 7.2 ± 1.6 years with classic CAH due to 21-hydroxylase deficiency treated with hydrocortisone and fludrocortisone. Multivariate linear mixed effects model analysis revealed a positive influence of CAH-specific z scores of summed urinary androgen metabolites (B = 0.97 ± 0.20, t = 4.87, P < 0.0001) and a negative influence of the cortisol metabolite tetrahydrocortisol (B = -1.75 ± 0.79, t = -2.20, P = 0.03) on HV z scores. Receiver operating characteristic analysis demonstrated that adrenal androgen excess, defined as HV >1.5 z, was best determined by a z score of all urinary androgen metabolites of >0.512 [accuracy, 66.2%; sensitivity, 57.1%; specificity, 74.4%; positive prediction value (PPV), 66.7%; negative prediction value (NPV), 65.9%]. Tetrahydrocortisol excretion >1480 µg/m2 BSA/d in conjunction with suppressed urinary androgen metabolites <0.163 z indicated overtreatment, defined as HV < -1.5 z (accuracy, 79.6%; sensitivity, 40.0%; specificity, 94.9%; PPV, 75.0%; NPV, 80.4%). We established target values for urinary steroid metabolite excretions in children with CAH based on their growth rate. Urinary steroid metabolome analysis represents a highly suitable method for monitoring metabolic control in children with CAH.

THE PHYSICAL DEVELOPMENT OF RURAL SCHOOLCHILDREN WHO ARE BROUGHT UP IN THE FAMILY AND IN THE BOARDING SCHOOL

There was made an assessment of the physical development of 319 rural schoolchildren who are brought up in families and in boarding schools. Anthropometric data of boys and girls aged 6-17 of years were analyzed. Normal growth rates for 91.3-94.3% of rural schoolchildren were established; Tall children were more often identified in families, and undersized one were revealed among those living in boarding schools (up to 8.9%). Normal values of body mass index were determined in 69.7-88% of schoolchildren; Overweight - in 10-26,9%, and body mass deficit - in 1,9-3,4% of children. In children living in families, high values of body mass index were detected by the order more often than low values (21.9-26.9% and 1.9-3.4%). In boarding students, the body mass index was the same in all age periods. Normal ratios of waist circumference to hip circumference were determined in 66.7-90% of schoolchildren. In this case, the predominant deposition of fat on the hips was found only in boys (25.7-33.3%); Abdominal fat deposits were found in 2.9% of boys living in families and in 10-17.6% of girls living in families and boarding schools. The authors believe that irrespective of the living conditions (family or boarding school), rural schoolchildren develop changes in the distribution of fat, possibly due to the peculiarities of the pubertal period.

Crystal-plane-dependent effects of antifreeze glycoprotein impurity for ice growth dynamics.

An impurity effect on ice crystal growth in supercooled water is an important subject in relation to ice crystal formation in various conditions in the Earth's cryosphere regions. In this review, we consider antifreeze glycoprotein molecules as an impurity. These molecules are well known as functional molecules for controlling ice crystal growth by their adsorption on growing ice/water interfaces. Experiments on free growth of ice crystals in supercooled water containing an antifreeze protein were conducted on the ground and in the International Space Station, and the normal growth rates for the main crystallographic faces of ice, namely, basal and prismatic faces, were precisely measured as functions of growth conditions and time. The crystal-plane-dependent functions of AFGP molecules for ice crystal growth were clearly shown. Based on the magnitude relationship for normal growth rates among basal, prismatic and pyramidal faces, we explain the formation of a dodecahedral external shape of an ice crystal in relation to the key principle governing the growth of polyhedral crystals. Finally, we emphasize that the crystal-plane dependence of the function of antifreeze proteins on ice crystal growth relates to the freezing prevention of living organisms in sub-zero temperature conditions. This article is part of the theme issue 'The physics and chemistry of ice: scaffolding across scales, from the viability of life to the formation of planets'.

Improvement of nitrogen use efficiency in maize using molecular and physiological approaches

The germination percentages for all maize (10 inbred lines) used under N+ or N- nutritional regimes was determined. Significant germination differences for inbred 6, 8 and 10 were indicated, while the other differences were non-significant. A large amount of phenotypic diversity is observed across maize lines, attributed mostly to significant differences in genome structural and gene presence/absence variation. Inbred line number 10 seems to be the most homogeneous genotype among the lines tested. This line showed the most normal germination and growth rate under (N+) and (N-) regimes. During the second season, these two lines (10 and 8) showed normal germination and growth rate under (N+) and (N-) regimes. Both inbred lines were significant for leaf Nitrogen content while, line 10 showed more significant difference for leaf Nitrogen content under (N+) and (N-) nutritional regimes. In order to characterize some genes related to NUE, the differential display PCR was applied for leaf extract samples of inbred lines 8 and 10 at three different developmental stages 5, 7 and 10 weeks grown under N+ (T1, T2, and T3) and N- (C1, C2 and C3) regimes, respectively. Some of the specific NUE amplified fragments were expressed in two developmental stages. Therefore, from the total of 39 amplified fragments, the sum of 31 expressed genes was identified through our present work. The phylogenetic analysis showed that sequences of glutamine synthetase gene are grouped among two clusters.

Milk biosynthesis requires the Golgi cation exchanger TMEM165

Milk is a hallmark of mammals that is critical for normal growth and development of offspring. During biosynthesis of lactose in the Golgi complex, H+ is produced as a by-product, and there is no known mechanism for maintaining luminal pH within the physiological range. Here, using conditional, tissue-specific knockout mice, immunostaining, and biochemical assays, we test whether the putative H+/Ca2+/Mn2+ exchanger known as TMEM165 (transmembrane protein 165) participates in normal milk production. We find TMEM165 is crucial in the lactating mammary gland for normal biosynthesis of lactose and for normal growth rates of nursing pups. The milk of TMEM165-deficient mice contained elevated concentrations of fat, protein, iron, and zinc, which are likely caused by decreased osmosis-mediated dilution of the milk caused by the decreased biosynthesis of lactose. When normalized to total protein levels, only calcium and manganese levels were significantly lower in the milk from TMEM165-deficient dams than control dams. These findings suggest that TMEM165 supplies Ca2+ and Mn2+ to the Golgi complex in exchange for H+ to sustain the functions of lactose synthase and potentially other glycosyl-transferases. Our findings highlight the importance of cation and pH homeostasis in the Golgi complex of professional secretory cells and the critical role of TMEM165 in this process.

Effect of Abiotic Stresses on Brassica Species and Role of Transgenic Breeding for Adaptation

Abiotic stresses extremely affect crop productivity and agro-morphological and biochemical properties of all Brassica species. Due to these stresses, yield of many important Brassica species decreases by disturbing their normal growth rate. In this review, we have highlighted the latest reports about the impact of different abiotic stresses on different growth stages and other morpho-physiological processes of important Brassica species such as canola/rapeseed (Brassica napus), Indian mustard (Brassica juncea), Brassica oleracea and Brassica rapa. Many researchers reported that abiotic stresses influence the important morpho-biochemical characteristics such as root and shoot length, shoot fresh and dry weight, proline and relative water contents, chlorophyll amount, antioxidant enzymes activity of important Brassica species. Cell injury also occurs by disturbance in normal oxidative processes due to these stresses. Against these stresses genetic modification have been described for the development of transgenic plant. The present study will be useful to identify the best abiotic stress tolerant Brassica genotypes for further genetic engineering program and crop improvement programs.

Abstract P2-06-19: The effect of UBE2C expression on intrinsic chemosensitivity in breast cancer cell lines

Abstract Background: The ubiquitin-proteasome pathway plays a crucial role in cancer-related processes by inducing cell cycle arrest through the degradation of mitotic cyclins and other cell cycle regulatory proteins. We recently showed that elevated levels of ubiquitin-conjugating enzyme E2C (UBE2C) were associated with aggressive tumor features and unfavorable clinical outcome in breast carcinoma (BC). UBE2C suppression has been achieved using the FDA-approved proteasome inhibitor bortezomib (VELCADE®) in colorectal carcinoma, but little is known about the efficacy of UBE2C-targeted therapy with proteasome inhibitors in breast cancer. Methods: Cell viability assays were used to determine the intrinsic chemosensitivity of five BC cell lines (MCF-7, MDA-MB-436, HCC38, HCC1395, and ZR-75-30; stratified by UBE2C expression and ER status) and the MCF-10A epithelial cell line to proteasome inhibitors (n=8), mitosis inhibitors (n=2), and platinum agents (n=3). UBE2C expression analysis was performed using quantitative real-time PCR and Western blot. IC50 values and growth inhibition metrics (GR50 and GRmax) were calculated for each compound to determine drug potency and efficiency after 24 hour treatment. Proteasome activity was assessed using bortezomib-treated cells. Results: Heterogeneous UBE2C expression levels were found in the different cell lines, with higher UBE2C levels in ER-negative BC cell lines (HCC38, HCC1395, MDA-MB-436) than ER-positive BC (MCF-7 and ZR-75-30) and MCF-10A control cells (ER-negative). Proteasome inhibition levels close to 50% and 100% were seen in all cell lines after 10 nM and 100 - 1000 nM bortezomib, respectively. As expected, bortezomib blocked cell cycle progression by inducing G2/M phase arrest in HCC38 cells. Due to differences in cell growth rates, calculation of the IC50 value was an ineffective method to determine drug potency. In contrast, the normalized growth rate inhibition method with GR50 and GRmax values demonstrated a correlation between sensitivity to proteasome inhibitors in ER-negative BC cell lines and high UBE2C expression levels. However, MDA-MB-436 cells (GR50, range 1.8-286.1 nM; GRmax, range -0.42- -0.93) were generally less sensitive to proteasome inhibitors than HCC38 cells (GR50, range 8.2-936.8 nM; GRmax, range -0.97- -0.99) though both cell lines were ER-negative, which was possibly due to the lower expression of UBE2C in MDA-MB-436 cells. Compared with the other tested drugs, no cell line was sensitive to the mitosis inhibitors and platinum agents were most effective in HCC38 cells. Conclusions: Taken together, these findings suggest an association between UBE2C expression and response to proteasome inhibition, regardless of ER status. Citation Format: Parris TZ, Larsson P, Biermann J, Engqvist H, Werner-Rönnerman E, Kovács A, Karlsson P, Helou K. The effect of UBE2C expression on intrinsic chemosensitivity in breast cancer cell lines [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P2-06-19.

Crystallization process and growth mechanism for hexagonal prism of strontium hydroxyapatite by urea hydrolysis

Strontium hydroxyapatite (SrHAp) is not only used as drug carrier material for bone disease treatment, but also has application prospect in bone tissue engineering. But SrHAp is prepared with by-products strontium hydrogen phosphate (SrHP) and strontium phosphate (TSrP), which requires accurate process optimizations and adjustments to circumvent the undesirable morphology products. The integral hexagonal prism crystals of SrHAp were successfully synthesized by urea hydrolysis at ambient pressure and without any strong acids and bases. Similarly, variable by-products are classified and investigated for their effects on crystallization process of strontium hydroxyapatite via monitoring the urea hydrolysis time-dependent products. The crystallization process of SrHAp can be divided into four stages based on the crystallographic results for the samples acquired by XRD and SEM studies during the entire synthesis processes. In the first stage, SrHP is formed (0–0.17 h); secondly, SrHP is converted into SrHAp (0.17–4 h); TSrP is obtained (4–8 h) at the third stage; and at last, TSrP is transformed into SrHAp (8–24 h), especially the amount of SrHAp obtained at 24 h was the sum of the second and fourth stages. Simultaneously, the growth mechanism of SrHAp crystals was proposed, considered of the formation energy of Gibbs-Thomson crystal plane and Born-Lande lattice energy. By calculating the interfacial energy γ(h k l) of different crystal planes, the γ(1 0 0) of hexagonal prism plane family m{1 0 0} is 1.3 × 10−5U0, the γ(0 0 1) of parallel double-plane family c{0 0 1} is 7.5 × 10−6U0, the normal growth rate of Ic{0 0 1} ≫ Im{1 0 0}. Accordingly, the growth mechanism for the preferential c axis growth maintaining the hexagonal prism shape is derived.