Life Sciences Commons^™

Linking Migration To Community Resilience In The Receiving Basin Of A Large-Scale Water Transfer Project, Anna Erwin, Zhao Ma, Ruxandra Popovici, Emma Patricia Salas O’Brien, Laura Zanotti, Chelsea A. Silva, Eliseo Zeballos Zeballos, Jonathan Bauchet, Nelly Ramírez Calderón, Glenn Roberto Arce Larreah

School of Earth, Environmental, and Marine Sciences Faculty Publications and Presentations

Large-scale water transfer projects (LWTPs) transfer water to urban and agricultural areas. The Majes-Siguas canal, established in 1983, is an LWTP that created a thriving agricultural area through irrigating the Majes district in the Atacama Desert of Peru. Like other LWTP receiving basins, the project has attracted an influx of migrants who work on the farms. At the same time, the Majes LWTP is the district’s only source of water and has an aging infrastructure which presents significant risks. While many studies critically analyze the consequences of LWTPs in water supply basins, few evaluate the resilience of communities living in …

Late-Life Exercise Mitigates Skeletal Muscle Epigenetic Aging, Kevin A. Murach, Andrea L. Dimet-Wiley, Yuan Wen, Camille R. Brightwell, Christine M. Latham, Cory M. Dungan, Christopher S. Fry, Stanley J. Watowich

Center for Muscle Biology Faculty Publications

There are functional benefits to exercise in muscle, even when performed late in life, but the contributions of epigenetic factors to late-life exercise adaptation are poorly defined. Using reduced representation bisulfite sequencing (RRBS), ribosomal DNA (rDNA) and mitochondrial-specific examination of methylation, targeted high-resolution methylation analysis, and DNAge™ epigenetic aging clock analysis with a translatable model of voluntary murine endurance/resistance exercise training (progressive weighted wheel running, PoWeR), we provide evidence that exercise may mitigate epigenetic aging in skeletal muscle. Late-life PoWeR from 22–24 months of age modestly but significantly attenuates an age-associated shift toward promoter hypermethylation. The epigenetic age of muscle …

Sap Beetles, Nick Volesky, Zachery R. Schrumm, Kalen Taylor

All Current Publications

Sap beetles are typically considered a secondary pest of corn and overripe fruits and vegetables. They are broadly identified by their small and ovular bodies and club-shaped antennae. Adult sap beetles often feed on corn silk, pollen, and tassels. Larvae feed on kernels inside the husk. This fact sheet reviews how to identify sap beetles and their hosts and damage. It also provides ideas on how to monitor for sap beetles and outlines the options for managing them.

Induction Of Il19 Expression Through Jnk And Cgas-Sting Modulates Dna Damage–Induced Cytokine Production, Sara H. Small, E. Jessica Tang, Ryan L. Ragland, Yaroslava Ruzankina, David W. Schoppy, Rahul S. Mandal, M. Rebecca Glineburg, Zgjim Ustelenca, Daniel J. Powell, Fiona Simpkins, F. Bradley Johnson, Eric J. Brown

Biology, Chemistry, and Environmental Sciences Faculty Articles and Research

Cytokine production is a critical component of cell-extrinsic responses to DNA damage and cellular senescence. Here, we demonstrated that expression of the gene encoding interleukin-19 (IL-19) was enhanced by DNA damage through pathways mediated by c-Jun amino-terminal kinase (JNK) and cGAS-STING and that IL19 expression was required for the subsequent production of the cytokines IL-1, IL-6, and IL-8. IL19 expression was stimulated by diverse cellular stresses, including inhibition of the DNA replication checkpoint kinase ATR (ataxia telangiectasia and Rad3-related protein), oncogene expression, replicative exhaustion, oxidative stress, and DNA double-strand breaks. Unlike the production of IL-6 and IL-8, IL19 expression was …

Cryptococcus Neoformans Melanization Incorporates Multiple Catecholamines To Produce Polytypic Melanin, Rosanna P. Baker, Christine Chrissian, Ruth E. Stark, Arturo Casadevall

Publications and Research

Melanin is a major virulence factor in pathogenic fungi that enhances the ability of fungal cells to resist immune clearance. Cryptococcus neoformans is an important human pathogenic fungus that synthesizes melanin from exogenous tissue catecholamine precursors during infection, but the type of melanin made in cryptococcal meningoencephalitis is unknown. We analyzed the efficacy of various catecholamines found in brain tissue in supporting melanization using animal brain tissue and synthetic catecholamine mixtures reflecting brain tissue proportions. Solid-state NMR spectra of the melanin pigment produced from such mixtures yielded more melanin than expected if only the preferred constituent dopamine had been incorporated, …

Ecological Risk Assessment For The State-Wide Small Pelagic Scalefish Resource, S. Blazeski, J. Norriss, K. A. Smith, M. Hourston

Fisheries research reports

In July 2021, the Department of Primary Industries and Regional Development convened an ecological risk assessment (ERA) of the fisheries that access the State-wide Small Pelagic Scalefish Resource. The ERA considered the potential ecological impacts of the West Coast Purse Seine Fishery, South Coast Purse Seine Fishery, Purse Seine Development Zones and the recreational fishers who catch small pelagic scalefish. The assessment evaluated the impact of each fishing sector/method on all relevant retained and bycatch species, endangered, threatened and protected species, habitats and the broader environment.

Domestication Reshaped The Genetic Basis Of Inbreeding Depression In A Maize Landrace Compared To Its Wild Relative, Teosinte, Luis Fernando Samayoa, Bode A. Olukolu, Chin Jian Yang, Qiuyue Chen, Markus G. Stetter, Alessandra M. York, Jose De Jesus Sanchez-Gonzalez, Jeffrey C. Glaubitz, Peter J. Bradbury, Maria Cinta Romay, Qi Sun, Jinliang Yang, Jeffrey Ross-Ibarra, Edward S. Buckler, John F. Doebley, James B. Holland

Department of Agronomy and Horticulture: Faculty Publications

Inbreeding depression is the reduction in fitness and vigor resulting from mating of close relatives observed in many plant and animal species. The extent to which the genetic load of mutations contributing to inbreeding depression is due to large-effect mutations versus variants with very small individual effects is unknown and may be affected by population history. We compared the effects of outcrossing and self-fertilization on 18 traits in a landrace population of maize, which underwent a population bottleneck during domestication, and a neighboring population of its wild relative teosinte. Inbreeding depression was greater in maize than teosinte for 15 of …

Identification Of Candidate Genes And Genomic Regions Associated With Adult Plant Resistance To Stripe Rust In Spring Wheat, Amira M. I. Mourad, Mohamed A. Abou-Zeid, Shamseldeen Eltaher, P. Stephen Baenziger, Andreas Börner

Department of Agronomy and Horticulture: Faculty Publications

Wheat stripe rust (caused by Puccinia striiformis f. sp. tritici) is a major disease that damages wheat plants and affects wheat yield all over the world. In recent years, stripe rust became a major problem that affects wheat yield in Egypt. New races appeared and caused breakdowns in the resistant genotypes. To improve resistance in the Egyptian genotypes, new sources of resistance are urgently needed. In the recent research, a set of 95 wheat genotypes collected from 19 countries, including Egypt, were evaluated for their resistance against the Egyptian race(s) of stripe rust under field conditions in the two …

Stimulation Of Ammonia Oxidizer And Denitrifier Abundances By Nitrogen Loading: Poor Predictability For Increased Soil N2o Emission, Yong Zhang, Feng Zhang, Diego Abalos, Yiqi Luo, Dafeng Hui, Bruce A. Hungate, Pablo García-Palacios, Yakov Kuzyakov, Jørgen Eivind Olesen, Uffe Jørgensen, Ji Chen

Biology Faculty Research

Unprecedented nitrogen (N) inputs into terrestrial ecosystems have profoundly altered soil N cycling. Ammonia oxidizers and denitrifiers are the main producers of nitrous oxide (N₂O), but it remains unclear how ammonia oxidizer and denitrifier abundances will respond to N loading and whether their responses can predict N-induced changes in soil N₂O emission. By synthesizing 101 field studies worldwide, we showed that N loading significantly increased ammonia oxidizer abundance by 107% and denitrifier abundance by 45%. The increases in both ammonia oxidizer and denitrifier abundances were primarily explained by N loading form, and more specifically, organic N …

A Cross-Ecoregion Evaluation Of Nitrogen Fixation And Denitrification In Streams And Rivers Of The United States Of America, Amy Marcarelli, Erin K. Eberhard, Michelle Kelly, Kevin Nevorski

Michigan Tech Publications

It is typically assumed that dinitrogen (N2) fixation and denitrification are mutually exclusive processes in riverine ecosystems because N2 fixation is favored in high light, low nitrogen (N) environments but denitrification is favored under anoxic, high N conditions. Yet recent work in marine and lake ecosystems has demonstrated that N2 fixation can happen under high N conditions and in sediments, challenging this assumption. We conducted a cross-ecoregion study to test the hypothesis that N2 fixation and denitrification would co-occur in streams and rivers across a range of reactive N concentrations. Between 2017 and 2019, we sampled 30 streams in 13 …

Β-Hydroxy-Β-Methylbutyrate Supplementation Promotes Antitumor Immunity In An Obesity Responsive Mouse Model Of Pancreatic Ductal Adenocarcinoma, Michael F. Coleman, Kristyn A. Liu, Alexander J. Pfeil, Suhas K. Etigunta, Xiaohu Tang, Salvador Fabela, Et. Al.

Michigan Tech Publications

Pancreatic cancer (PDAC) is a deadly disease, exacerbated by obesity, which lacks effective therapeutic interventions. Most PDAC has a limited response to immune- and chemotherapy. Treating PDAC is made additionally challenging by the rapid emergence of muscle wasting and cachexia, which predict poor response to several therapies. We have found that dietary supplementation with β-hydroxy-β-methylbutyrate promotes immunosurveillance in PDAC tumors and protects muscle. This dietary supplement has the potential to be an important adjuvant in PDAC therapy, opening the doors to immunotherapy response.

Reaction Of Alkynyl- And Alkenyltrifluoroborates With Propargyldicobalt Cations: Alkynylation, Alkenylation, And Cyclopropanation Product Pathways, Brent St Onge, S. Maryamdokht Taimoory, Jeffrey Battersby, John F. Trant, James R. Green

Chemistry and Biochemistry Publications

The Lewis acid-mediated Nicholas reactions of propargyl acetate–Co2(CO)6 complexes with a series of potassium alkynyltrifluoroborates and potassium alkenyltrifluoroborates are described. Alkynyltrifluoroborates directly alkynylate the intermediate propargyldicobalt cations. In contrast, alkenyltrifluoroborates proceed through one of the three modes of dominant reactivity: C-2-substituted alkenyltrifluorobrates directly alkenylate, predominantly with the retention of stereochemistry. C-1-substituted alkenyltrifluoroborates alkenylate at C-2. Potassium vinyltrifluoroborate incorporates a cyclopropane at the site propargyl to alkynedicobalt. Computational analysis of these systems explains the differential modes of reactivity of alkenyltrifluoroborates and outlines the probable mechanisms for the formation of each product.

Protocol For Fabricating Electroless Nickel Immersion Gold Strain Sensors On Nitrile Butadiene Rubber Gloves For Wearable Electronics, Sara S. Mechael, Yunyun Wu, Yiting Chen, Tricia Breen Carmichael

Chemistry and Biochemistry Publications

This protocol describes the fabrication of patterned conductive gold films on nitrile butadiene rubber (NBR) gloves for wearable strain sensors using electroless nickel immersion gold (ENIG) plating, a solution-based metallization technique. The resulting NBR/ENIG films are strain sensitive; resistance measurements of a patterned sensing array can be used to map human hand motions. This protocol also describes challenges related to the ENIG process and troubleshooting steps to achieve conformal gold films for strain sensing over a large working range. For complete details on the use and execution of this protocol, please refer to Mechael et al. (2021).

A Protocol For Rapid And Parallel Isolation Of Myocytes And Non-Myocytes From Multiple Mouse Hearts., Gabriella E Farrugia, Michael A Mclellan, Kate L Weeks, Aya Matsumoto, Charles D Cohen, Crisdion Krstevski, Taylah L Gaynor, Adam C Parslow, Julie R Mcmullen, Alexander R Pinto

Faculty Research 2021

This protocol features parallel isolation of myocytes and non-myocytes from murine hearts. It was designed with considerations for (1) time required to extract cardiac cells, (2) cell viability, and (3) protocol scalability. Here, a peristaltic pump and 3D-printed elements are combined to perfuse the heart with enzymes to dissociate cells. Myocytes and non-myocytes extracted using this protocol are separated by centrifugation and/or fluorescence-activated cell sorting for use in downstream applications including single-cell omics or other bio-molecular analyses. For complete details on the use and execution of this protocol, please refer to McLellan et al. (2020).

Farnesol Secretion As A Possible Driving Force For Maintaining Candida Albicans As A Diploid, Kenneth Nickerson, Cory Boone, Kory Parker

Kenneth Nickerson Papers

Candida albicans is a pathogenic dimorphic fungus which is invariably found as a diploid in patients. C. albicans secretes the sesquiterpene farnesol both as a quorum sensing molecule which blocks the yeast to hypha conversion and as a virulence factor for pathogenicity. 20-25 μM farnesol kills other competing yeasts and fungi, often by triggering apoptosis, and yet wild type diploid C. albicans tolerates 300-500 μM farnesol. The recent availability of 10 haploid strains of C. albicans (5 mating type aand 5 mating type α) allowed us to compare their production of and sensitivity to farnesol. On average, the heterozygous diploid …

Relationships Among College-Level Science Course Enrollment, Environmental Perception, And Pro-Environmental Attitude: Evidence From The Us General Social Survey, Mazbahul G. Ahamad, Fahian Tanin

Department of Agronomy and Horticulture: Faculty Publications

Understanding pro-environmental attitudes is critical to encouraging pollution-minimizing behaviors. Therefore, identifying associated factors is essential for understanding different types of pro-environmental attitudes. We aimed to investigate the associations among individuals’ college-level science course enrollment and their perceptions of the level of spending to improve and protect the environment, as well as their pro-environmental attitudes. We used nationwide population-based cross-sectional survey data from 2,348 individuals obtained from the General Social Survey in the United States. An ordered logistic model was used to examine the associations among college-level science course enrollment, environmental perception, and pro-environmental attitude. We found that science course enrollment …

S5e11: Why Might Maine Lose Two Species Of Songbirds?, Ron Lisnet, Katherine Ruskin, Brian Olsen

The Maine Question

Maine may lose two tidal marsh songbird species in the next few decades. Saltmarsh sparrows face extinction, while Acadian Nelson’s sparrows are threatened with extirpation — localized eradication with the possibility of survival elsewhere. Their populations along the Eastern Seaboard have been declining as sea level rise destroys their habitats and, according to a new University of Maine-led study, mercury exposure inhibits their reproduction.

In the final episode of Season 5 of “The Maine Question,” Kate Ruskin, a lecturer in ecology and environmental science at UMaine who spearheaded the recent mercury exposure study, and Brian Olsen, who is now an …

Population Ecology Of The Queensnake (Regina Septemvittata) In An Urban Creek, 2008 To 2019, Rachel Beiler, R. Elliot Miller, Norman Reichenbach

Faculty Publications and Presentations

Habitat fragmentation is a common result of urbanization and species living in these fragments are at risk of extirpation. We conducted a 12-y (2008–2019) capture-mark-recapture study on snakes living in a 593-m section of Rock Castle Creek flowing through an urban area in central Virginia, USA. Our study site was occupied primarily by Queensnakes (Regina septemvittata). We used data from 168 individual Queensnakes to examine several aspects of their population ecology including survival rates (0.52), reproductive effort (21.3% juvenile to adult ratio), growth rates (68% and 30.6% increase for 1 to 2 y-old and 2 to 3+ y-old snakes, respectively), …

Characterization Of Landfill Leachate For Enhanced Metal Recovery, Hanna Fulford, Amisha Shah, Inez Hua, Nadezhda Zyaykina, Lori Hoagland, Alejandro Rodriguez Sanchez, Umut Bicim

Discovery Undergraduate Interdisciplinary Research Internship

Landfills contain a trove of valuable materials, such as critical, precious, and rare earth metals, that are integral to the United State’s economy and national security. The leachate that filters through landfills picks up these materials, which allows for the possibility of recovery. For this research, samples will be analyzed from landfills throughout the Midwestern United States to provide a baseline on water quality constituents, elements present, and microbial activity. Preliminary data for this study was acquired by analyzing samples of landfill leachate from a landfill in northern Indiana. pH readings indicate that the leachate is slightly basic. It also …

Skin Bacterial Metacommunities Of San Francisco Bay Area Salamanders Are Structured By Host Genus And Habitat Quality., Shannon Buttimer, Obed Hernandez-Gomez, Erica Bree Rosenblum

Natural Sciences and Mathematics | Faculty Scholarship

Host-associated microbial communities can influence physiological processes of macroorganisms, including contributing to infectious disease resistance. For instance, some bacteria that live on amphibian skin produce antifungal compounds that inhibit two lethal fungal pathogens, Batrachochytrium dendrobatidis (Bd) and Batrachochytrium salamandrivorans (Bsal). Therefore, differences in microbiome composition among host species or populations within a species can contribute to variation in susceptibility to Bd/Bsal. This study applies 16S rRNA sequencing to characterize the skin bacterial microbiomes of three widespread terrestrial salamander genera native to the western United States. Using a metacommunity structure analysis, we identified dispersal barriers for these influential bacteria between salamander …

Restoring Polyhyrdoxybutyrate (Phb) Depolymerase Expression In A Bald Mutant Of Streptomyces Sp. Sfb5a, Youmna Moawad

Honors Projects

Polyhydroxybutyrate (PHB) is a biodegradable, plastic-like polymer produced by some bacteria and degraded by others. The filamentous bacterium Streptomyces sp. SFB5A degrades PHB for growth using an extracellular PHB depolymerase, encoded by the phaZ gene. A morphological mutant of this bacterium, bld4, cannot form aerial filaments and cannot produce PHB depolymerase, despite having the phaZ gene. The inability to synthesize PHB depolymerase may be due to a mutation in a gene encoding one of its transcriptional regulators. A gene (lrp) coding for a potential transcriptional regulator is located 2,700 base pairs upstream from phaZ. Our goal …

Genetic Diversity Of Huaya India (Melicoccus Oliviformis Kunth), A Neglected Neotropical Fruit Crop, Mónica I. Jiménez-Rojas, Rubén H. Andueza-Noh, Obed I. Noh-Ake, Daniel Potter, Matilde M. Ortiz-García, Renee S. Arias, Jaime Martínez-Castillo

United States Department of Agriculture-Agricultural Research Service / University of Nebraska-Lincoln: Faculty Publications

Currently, some species of Sapindaceae are important fruit crops worldwide. The Huaya India (Meliccocus oliviformis, Sapindaceae) is a neglected Neotropical fruit tree consumed locally in the Maya Lowlands of Mexico, where it exists in both wild and domesticated forms. Our objective was to evaluate the genetic diversity of the Huaya India in its possible domestication area and thus generate knowledge that serves as the basis for a commercial management. A total of 450 individuals collected from 15 natural vegetation sites and 15 Maya villages, were characterized using nine microsatellite loci and population genetics approaches were applied. STRUCTURE, Neighbor-Joining and PCoA …

Finland, A Package Deal: Disciplinary Climate In Science Classes, Science Dispositions And Science Literacy, Larry J. Grabau, Jari Levonen, Kalle Juuti

Plant and Soil Sciences Faculty Publications

Finland’s educational prowess, though tempered by recent international assessments, has remained intact. This report focused on lessons that could be learned regarding secondary-level science education from the Program for International Student Assessment (PISA) 2015, science-focused assessment. That PISA iteration included not only science literacy but also students’ science dispositions (epistemology, enjoyment, interest, and self-efficacy) and the schools’ science climate measures (disciplinary climate and teaching support). Due to the hierarchical nature of the PISA data, multilevel models were employed in this Finnish study, involving 5582 students from 167 schools. Science dispositions (as outcome measures) were differently associated with teaching support and …

Greta Thunberg: A Small But Mighty Voice For The Environment, Madilyn Mortelliti

Faculty Curated Undergraduate Works

Abstract

This research paper discusses Greta Thunberg and the impact she has as a climate change activist. Thunberg faces many challenges as a young woman, but overcomes these difficulties while encouraging others to follow in her footsteps. She presents herself as a positive role model for younger generations. Thunberg has many accomplishments as an extremely young woman that leads to her global iconicity. She makes her power known in front of the highest ranked men in the world which forces legislation to make a change. Thunberg motivates others to make a difference before all hope is lost.

Measuring Science Teachers' Emotional Experiences With Evolution Using Real World Scenarios, William Lee Romine, Rutuja Mahajan, Amber Todd

Biological Sciences Faculty Publications

Low acceptance of evolution remains an obstacle to quality biology instruction. We develop and utilize a novel assessment which measures emotional experience in light of real-world evolution education scenarios. We presented 296 science teachers 4 pro-evolution and 8 anti-evolution scenarios and asked them to rate their levels of joy, anger, sadness, fear, disgust, shame, and guilt elicited by that scenario on an ordinal 5-point scale. We used exploratory factor analysis to extract the most important dimensions in the teachers’ responses, Rasch analysis to explore the validity of the extracted subscales, and stepwise regression to find the most important factors driving …

Bases For The Establishment Of Robusta Coffee (Coffea Canephora) As A New Crop For Colombia, Luis F. Campuzano-Duque, Juan Carlos Herrera, Claire Ged, Matthew Wohlgemuth Blair

Agricultural and Environmental Sciences Faculty Research

Robusta coffee (C. canephora) covers 36% of world coffee production and has strategic relevance as a beverage that it is produced by thousands of small-scale producers around the world. Although mainly grown in Africa and Asia as opposed to Latin America, this situation is changing. Colombia is recognized as a producer of high-quality Arabica (C. arabica L.) coffee, however we argue that Robusta represents a great economic opportunity for small scale producers, for the industrialization of new products and for emerging coffee chains. Therefore, the objective of this review is to outline the agronomic value of Robusta coffee as a …

Driver Liability For Farm And Ranch Operations, Jessica Groskopf

Cornhusker Economics

One major concern of farming and/or ranching operations is the liability of semi-trucks and equipment when they are involved in a traffic accident.

Cold Conditioned: Discovery Of Novel Alleles For Low-Temperature Tolerance In The Vavilov Barley Collection, Ahmad H. Sallam, Kevin P. Smith, Gongshe Hu, Jamie Sherman, Peter Stephen Baeziger, Jochum Wiersma, Carl Duley, Eric J. Stockinger, Mark E. Sorrells, Tamas Szinyei, Igor G. Loskutov, Olga N. Kovaleva, Jed Eberly, Brian J. Steffenson

Department of Agronomy and Horticulture: Faculty Publications

Climate changes leading to higher summer temperatures can adversely affect cool season crops like spring barley. In the Upper Midwest region of the United States, one option for escaping this stress factor is to plant winter or facultative type cultivars in the autumn and then harvest in early summer before the onset of high-temperature stress. However, the major challenge in breeding such cultivars is incorporating sufficient winter hardiness to survive the extremely low temperatures that commonly occur in this production region. To broaden the genetic base for winter hardiness in the University of Minnesota breeding program, 2,214 accessions from the …

Effect Of Biofertilizer In Organic And Conventional Systems On Growth, Yield And Baking Quality Of Hard Red Winter Wheat, Ammar Al-Zubade, Timothy D. Phillips, Mark A. Williams, Krista L. Jacobsen, David Van Sanford

Horticulture Faculty Publications

A two-year study (harvest years 2019 and 2020) was conducted to investigate the effect of a commercially available biofertilizer, in combination with variable nitrogen (N) rate, on bread baking quality and agronomic traits in hard winter wheat grown in conventional (CONV) and organic (ORG) farming systems in Kentucky, USA. The hard red winter wheat cultivar ‘Vision 45’ was used with three N rates (44, 89.6 and 134.5 kg/ha as Low, Med and High, respectively) and three biofertilizer spray regimes (no spray, one spray and two sprays). All traits measured were significantly affected by the agricultural production system (CONV or ORG) …

Spatial Variability In Streambed Microbial Community Structure Across Two Watersheds, Philips Akinwole, Jinjun Kan, Louis A. Kaplan, Robert H. Findlay

Biology Faculty publications

Both spatial and temporal variability are key attributes of sedimentary microbial communities, and while spatial effects on beta-diversity appear to dominate at larger distances, the character of spatial variability at finer scales remains poorly understood, especially for headwater stream communities. We investigated patterns of microbial community structure (MCS) in biofilms attached to streambed sediments from two watersheds across spatial scales spanning ,1 m within a single stream to several hundred kilometers between watersheds. Analyses of phospholipid fatty acid (PLFA) profiles indicated that the variations in MCS were driven by increases in the relative abundance of microeukaryotic photoautotrophs and their contribution …