*Article* **Electrospun Sericin/PNIPAM-Based Nano-Modified Cotton Fabric with Multi-Function Responsiveness**

**Jia Li 1,2,3, Bo-Xiang Wang 2,3, De-Hong Cheng 2,3, Zhi-Mei Liu 3, Li-Hua Lv 1, Jing Guo 1,4,\* and Yan-Hua Lu 2,3,\***

**Abstract:** There is a significant interest in developing environmentally responsive or stimuli-responsive smart materials. The purpose of this study was to investigate multi-function responsive cotton fabrics with surface modification on the nanoscale. Three technologies including electrospinning technology, interpenetrating polymer network technology, and cross-linking technology were applied to prepare the multi-function sericin/poly(N-isopropylacrylamide)/Poly(ethylene oxide) nanofibers, which were then grafted onto the surfaces of cotton textiles to endow the cotton textiles with outstanding stimuliresponsive functionalities. The multi-function responsive properties were evaluated via SEM, DSC, the pH-responsive swelling behavior test and contact angle measurements. The results demonstrate that with this method, multi-function responsive, including thermo- and pH-responsiveness, cotton fabrics were fast formed, and the stimuli-responsiveness of the materials was well controlled. In addition, the antimicrobial testing reveals efficient activity of cotton fabrics with the sericin/PNIPAM/PEO nanofiber treatments against Gram-positive bacteria and Gram-negative bacteria such as *Staphylococcus aureus* and *Escherichia coli*. The research shows that the presented strategy demonstrated the great potential of multi-function responsive cotton fabrics fabricated using our method.

**Keywords:** sericin; poly(N-isopropylacrylamide); cotton fabrics; electrospinning


**4. Implications for farmed and wild bivalves**

Coastal water quality is strictly correlated to the food safety of bivalves. In fact, mollusc bivalves are filter feeders, and they can accumulate microbiological and chemical contaminants from surrounding water. Bivalves are generally cultivated or collected along with the coastal areas where their nutrients are abundant and flow from inland water. But the rivers can also discharge faecal bacteria, mostly from untreated wastewater [80]. The malfunctioning of urban waste water treatment plants or their by-pass during the severe rain events can contribute to the release faecal bacteria into the river. This can pose a potential risk to the consumers of bivalves that can accumulate these bacteria.

To avoid any health human risks, according to the EU Regulation No 627/2019 [37] the competent authorities firstly classify the production areas (Class A, B, and C) through specific monitoring campaigns. Then, they continue to control the level of faecal contamination of the bivalves according to the specific surveillance monitoring plan. The bacterium *E. coli* is used as a faecal indicator organism. For the Class A assignment, for example, the samples (80%) shall not exceed 230 *E. coli* per 100 g of flesh and intravalvular liquid. From this Class, molluscs can be collected for direct human consumption. During the sanitary control, if the mollusc health standards are not met, the competent authorities shall close the production areas and/or reclassify them [37].

As a decision support system for the competent authority, several studies have investigated the correlation between the increase of bacterial concentration in molluscs and weather conditions [40–49, 81–83]. The prediction of local precipitation and river discharges have been used as early warning signals for mollusc bacterial contamination [84, 85]. The advantages are multiple—i) to avoid the collection of the potentially contaminated product; ii) to avoid any temporary closing of production areas; iii) to optimise the monitoring surveillance programme; iv) to ensure the health of consumers.

Generally, results demonstrated that the correlation is site-specific and it depends on numerous factors, such as the geographical location, land use, and catchment size.

From ancient times, the Adriatic basin is particularly devoted to the bivalve farming and fishing in the lagoons and along the coasts. Here, the influence of the weather condition and river run-off on the bivalve hygiene condition has been investigated [41, 86, 87].

In the central Adriatic coast of the Marche region, recently, Ciccarelli et al. [88] published the correlation between the concentration of *E. coli* in the natural banks of *Chamelea gallina* and the local precipitation from 2016 to 2020. The results showed that the rainfall events were significant for the increase of *E. coli* (> 230 MPN/100 g) in the molluscs collected from the south sampling points. In the same region, the increase of *Salmonella* spp. detected in bivalves was reconducted in 2015 and 2016 to the severe meteorological events [89].

In the Northern Adriatic Sea, the CADEAU project [90] developed specific indexes to evaluate the potential microbial pollution impact of urban waste water treatment plants on the farmed molluscs in the Municipality of Chioggia (Venice, Italy). It provides indexes of dilution for *E. coli* based on the bacterial decay due to salinity, temperature, and solar radiation [90].

In the following sections, we report some "site-specific" study cases carried out in the Abruzzo region, on the central Adriatic coast of Italy.

## **4.1 The study case of wild clams and farmed mussels in the Pescara province**

In 2021, Colaiuda et al. [41] published the case study in the Pescara province (Abruzzo Region, Italy) that was already detailed in the paragraph 2.2. Here, two production areas of wild clams (*C. gallina*) and one farm of mussels (*Mytilus galloprovincialis*) facing the Pescara River were investigated (**Figure 1**). In **Figure 1**, Pescara 1 and 2 are the production areas of clams, the other is the farm of mussels. Thanks to the CapRadNet project, this study executed a correlation analysis between river discharge trough to the CHyM model, precipitation in the catchment area, and the concentrations of *E. coli* detected in the bivalves during the official monitoring programme. The referring period was from August 1, 2015 to July 31, 2016. The EU reference method to detect *E. coli* was ISO-16649-3 [91]. Results were expressed as the most probable number – MPN per 100 g of flesh and intravalvular liquid of mollusc. Microbiological data were downloaded from the database of the project CAPS2 developed also the informative tool "CAPS2 WEB GIS" useful for the management of the production areas. The classification of the three production areas (Class A) was viewable in the CAPS2 WEB GIS [92]. The competent authority was the unique authorised user to modify the classification and the boundaries of the production areas in the CAPS2 WEB GIS.

The results showed that the concentration of *E. coli* in molluscs increased within 6 days of a river discharge peak (**Figure 2**). Moreover, 87% of cases of high concentration of *E. coli* were consequent to the increased river flow, while 60% of cases to the precipitation. These results suggested that the Pescara River discharge was the potential hydrometeorological driver of *E. coli* in facing molluscs to be further evaluated with specific sampling before and after discharge peak at the river mouth.

#### **4.2 The study case of mussel farm in the Teramo province**

The research project FORESHELL was funded by the FLAG Costa Blu through the 2014-20 EMFF programme of the Abruzzo Region. It is aimed at developing sanitary/weather-environmental predictive technological tools to enhance the efficiency and sustainability of a mussel farm in the Teramo province (Giulianova city, Abruzzo region, Italy) [93]. This production area of *M. galloprovincialis* was classified as Class A, and it is facing the Salinello and Vibrata Rivers far away almost 3 miles from the coast (**Figure 6**).

The hydrological model (CHyM) has analysed the hydrographic basins of the rivers and it has been forecasting the discharge peaks. Before and after these events, a sample of freshwater at the river mouths, and of molluscs and sea water at the farm have been collected for the *E. coli* detection [91]. Preliminary results showed that until September 2021, there were four meteorological events (**Table 1**) that did not cause a peak discharge at the river mouth. Results did not register a significant increase of *E. coli* in the mussels (**Figure 7**). At the same time, the environmental parameters such as sea water temperature, salinity, Chl-a, sea currents, and wave motion are acquired by the satellites and *in situ* probes.

*Coastal Water Quality: Hydrometeorological Impact of River Overflow and High-resolution… DOI: http://dx.doi.org/10.5772/intechopen.104524*

#### **Figure 6.**

*Sampling points at river mouths and at the farm in the Abruzzo region.*

**Table 1.**

*FORESHELL project: Description of meteorological events.*

The web application for data visualisation is under construction, as well as the early warning signalling to the farmer by mail/SMS/WhatsApp. The alerts are referred to the potential faecal contamination of molluscs predicted through hydrological data and other parameters that can damage the farm, such as high temperature and wave motion.

Furthermore, the growth of mussels is constantly monitored with biometric controls.

In conclusion, the first period of project execution was characterised by precipitation scarcity that did not cause any discharge peaks at the river mouths without any presence of *E. coli* in the molluscs. Further analyses are expected to be executed during the rainy period of the autumn and winter seasons.

#### **4.3 Satellite data for bivalves**

Recently, satellite data and maps are increasingly used for the identification of areas intended for aquaculture, for the knowledge of the environmental conditions useful for shellfish farming and fishing, for the prediction of potentially harmful events, etc. The knowledge of parameters such as temperature, salinity, and turbidity give important information to managing bivalve production. For example, data on Chl-a could be useful to understand the food disposal for molluscs or the prediction of algal bloom potential toxic.

The projects AQUACULTURE2000 and VALUE-SHELL analyse satellite data, such as pH, temperature, and CO2, to assess the possible contribution of mussel farming in sequestering carbon from seawater through the biocalcification processes in the northern Adriatic Sea by mitigating the effects of climate change [94].

In 2021, the total suspended matter, temperature, and Chl-a estimated from satellite acquisitions have been used to predict the presence of radioactivity in molluscs [95].

Along the Adriatic coast facing the Abruzzo region where several farms are placed, two pilot studies were conducted. The goal was to calibrate algorithm coefficients, at a local scale, to set up a processing chain that derives accurate concentration maps of chlorophyll and suspended solids from the satellite, as said, taking advantage of the high frequency of revisit time and high spatial resolution of the satellite acquisitions.

The first study estimated Chl-a and sediment dispersions in the sea, derived from Sentinel-2 images, compared with *in situ* data acquired by means of a multiparametric probe in the monitoring stations that Agenzia Regionale per la Tutela dell'Ambiente (ARTA) Abruzzo monthly checks [96]. The Case-2 Regional Coast Color processor in ESA SNAP software was used, applying the C2RCC-Nets algorithm [97], whose parameters have been set using *in situ* measurements, specifically the salinity and temperature variables. This preliminary study provided encouraging results with only four sampling dates—for example, the concentration map of TSM of 09/03/2018 is reported (**Figure 8**).

In the second study [98], the authors developed the Water Color Data Analysis System (WC-DAS), a tool for the operational generation of maps and indicators useful in the monitoring of water quality. The tool, in its first release, allows the processing of satellite optical multispectral data acquired by Sentinel-3 OLCI, Sentinel-2 MSI, and Landsat-8 OLI sensors, using the algorithms Case 2 Regional CoastColour (C2RCC) [97] and Atmospheric Correction for OLI "lite" (ACOLITE) [99].

The tool was tested in the central Adriatic coastal zone, setting the local parameters according to the *ad hoc in situ* sampling campaign that ARTA Abruzzo carried out along the Abruzzo coast, simultaneously with the satellites overpasses (example in **Figure 9**).

The results show performances of calibrated algorithms and the data system's suitability to contribute to the production of monitoring maps and indicators,

*Coastal Water Quality: Hydrometeorological Impact of River Overflow and High-resolution… DOI: http://dx.doi.org/10.5772/intechopen.104524*

informing domain-specific decision-making and supporting services for integrated coastal zone management.

#### **Figure 8.**

*Concentration maps of Total Suspended Matter along Abruzzo coast in the Adriatic Sea, as elaborated with the C2RCC processor from Sentinel-2 imagery of 09/03/2018.*

#### **Figure 9.**

*Turbidity, Total Suspended Matter, Chlorophyll-a maps along Abruzzo coast, facing Pescara river mouth, as derived from Water Colour Data Analysis System Tool.*


**6. The Anti-Politics of Wind Power Development**

In response to the popular opposition and legal challenge to their investment, the LTWP project used various mechanisms to circumvent claims of illegal dispossession of communities and irregularities in the process of EIA and community consultation. Despite a lack of strong grassroots resistance to the project, top-down counterresistance was manifested through the actions of government institutions and corporate actors seeking to counterbalance and contain the dominance of communities' claims in their material and symbolic terms. In the socioeconomic context of "political reaction from above", the state-led and private sectors delivered development visions constituting the centerpiece of legitimising dispossession. Here, we define political reactions from above as multipronged strategies and techniques that serve a "state-driven, productive, employment-generating form of national development that enjoyed significant ideological legitimacy" [68], with the aim of making LSLA operations politically and socially feasible. By their very nature, these strategies express a discontent with local livelihoods and related land uses through particular conceptions of viability embodied in technical recommendations on the "economic units" and based on modernisation narratives [7]. The proponents of wind power in Northern Kenya sought to circumvent claims of nonadherence to legal procedures for community land acquisition and the need for international instruments for free informed and prior consent (FPIC) of local indigenous communities. The strategies employed by the corporate actors in LTWP project are highlighted below.

#### (a) Discursive regimes of representation

The premise of development discourse by the Kenyan government and the corporate actors involved in the LTWP contained in the development blueprint, *the Kenya Vision 2030*, and extensively reproduced in the LEC, positions the development of the hitherto marginalised north, energy deficiency and visions of affordable green energy as the centerpiece, legitimising exploitation and dispossession which characterised the process of land acquisition for the project. Kenya's 2002 elections not only brought Moi's 24-year rule to a close but also ushered in an era of development visions that sought to identify the key policy actions and reforms with an objective of realising a "higher and sustainable growth of the economy in a more equitable environment, accompanied by increased employment opportunities [69]". Kenya's *Vision 2030* resonates with discourses of "frontier economies" and "Africa rising" [5] and a revolutionary character through "modern opportunities for creating wealth (President Uhuru Kenyatta during the ground breaking ceremony for the LTWP Project quoted in Danwatch (2016) [70])", involving "new mindsets and methods, innovative strategies [26]". At the same time, however, these visions derive from and are embedded in the institutional patronages and settings that frame them. These ensuing visions, in addition to recognizing the "obscured but untapped potential" of marginalised areas such as Northern Kenya, also present a construction based on neoliberal ideologies of economic development that seek to correct existing social and economic inequalities through priority investments in infrastructure, energy and land reform. Considering the

historical evolution of development frames in Kenya, the emergent processes are likely to be less profound and less far-reaching [5].

The framing of the development of Northern Kenya is influenced by "technology transfer approaches, in which 'scientific' farming practices and technologies are provided to help modernise and civilise 'backward' farming systems" [7] and specifically refers to frontier economies as emerging centers of opportunities and engines of growth [71]. In this context, the government discourse is taken up by the corporate actors and positioned as "putting the lands to better use" for the benefit of local communities as well as meeting national development goals. The LTWP attorney stated the following in court proceedings at the March 2016 sitting of the LEC in Meru:

"Mr. Nyaoga told the Court that the respondents and the interested parties recognised the importance of the challenged project. He said that it was a Vision 2030 project. He said that the community had benefited immensely and will continue to benefit (Mr. Nyaoga, advocate for LTWP project. Civil Suit 163 of 2014 (Formerly Nairobi ELC 1330 of 2014) | Kenya Law Reports 2018. Page 11 of 28. http://kenyalaw.org/caselaw/cases/view/130470. Accesses on 29 June 2018)."

"for the first time since independence, the interested parties were seeing the fruits of independence through the challenged project (Mr. Kiprop, advocate for interested parties. Civil Suit 163 of 2014 Kenya Law Reports 2018. Page 11 of 28. http://kenyalaw.org/caselaw/ cases/view/130470. Accesses on 29 June 2018)."

The imperative of generating more energy at lower cost to meet the increased demand for energy in the realisation of the *Kenya Vision 2030* as well as reducing energy costs in Kenya, which are higher than those of all regional competitors, is another argument often made to legitimise the project. This discourse supports the admission by Kenya's government of energy insufficiency and the role of more private sector players in the energy sector [69]. The LTWP's discourse on wind power development heavily aligns with energy reliability narratives of *Kenya Vision 2030*:

"Kenya has developed policies that align with these low emission development strategies (LEDS) outcomes. For example, *Kenya Vision 2030* includes a National Climate Change Action Plan highlighting the importance of developing a secure, climate-resilient national grid that can support Kenya's development ambitions. In addition, Kenya's Draft National Energy and Petroleum Policy (2015) aims to ensure an affordable, competitive, sustainable, and reliable supply of energy to meet national development needs at lowest cost, while protecting and conserving the environment."

"These goals are crucial for Kenya's development, as fewer than 50% of Kenya's population have access to electricity."

"LTWP constructed a 208 km road from Laisamis to Sarima, which has transformed the transportation network in the area. This has significantly increased access to markets as well as to health care and educational facilities for the local population (WOC (2018). Community projects. https://ltwp.co.ke/winds-of-change/)."

Moreover, the LTWP portrays wind power development within the green energy discourse validated against a host of standards and certification schemes and international environmental treaties. The project was registered as a Clean Development Mechanism (CDM) project with The United Nations Framework Convention on Climate Change (UNFCCC). Green energy fits within the image of Kenya as being among the countries pursuing low emission development strategies (LES)—countries that grow their economies while reducing GHG emissions [72]. Consequently, *Kenya Vision 2030* has a diverse strategy of generating more energy at a lower cost, including exploitation of new sources of renewable energy through legal and policy instruments that aim to ensure an affordable, competitive, sustainable, and reliable supply of energy to meet national development needs at the

lowest cost, including an emphasis on the need to reduce GHG emissions and encourage the use of renewable energy as a mitigation measure [69]. The LTWP understands and aligns their wind energy activities with this agenda while remaining extra confident of their investment gaining credibility and buy-in from a large cross section of stakeholders. The LTWP's discourse on green energy is clearly linked to the push and established targets for green energy and opportunities arising from their development:

"The electricity generated from LTWP will provide a cost-effective alternative to fossil fuels, decrease dependency on foreign fuel imports, and stabilise prices through reliable generation. Power from LTWP will be sold to the grid at about one-third of the cost of fossil fuel-generated electricity. Additionally, the capacity from LTWP is expected to reduce the country's reliance on fuel imports. These avoided fuel imports are anticipated to save Kenya more than €100 million (US\$113 million) per year, improving the country's energy security." [72]

This strategy of development visions and green energy discourse is very effective, with LEC noting that the matter before it "involves a massive project". It also involves "weighty issues", noting in addition that the "project is the single largest private investment in Kenya's history" and that "through the Power Purchase Agreement, public funds have been and continue to be utlised ( ... ) The fact that the project is massive appears not to be in dispute [57]". This not only creates favorable conditions for the continuation of the wind power projects despite the "alienation of such expansive piece of community land to a private entity", which amounts to "a contemporary land injustice" [57] but also such discourse depoliticises the wind power development by omitting equally weighty issues related to impacts of the project on local livelihoods, ancestral and cultural heritage and loss of key common pool resources.

#### (b) The anti-politics of commons governance

The depoliticisation of wind power development by the LTWP has been characterised by a planning process that casts local pastoral livelihoods and communal land use as "unviable" and a need to transform "idle" and "underutlised" land through an efficient infrastructure network, creating an enabling environment for private sector participation and harnessing wind, solar and geothermal energy among other strategies. This not only puts the responsibility of planning, constructing and operating large-scale development programs into the hands of private sector actors but also creates an illusion that state bureaucrats and local institutions are absolved from their responsibility and accountability [4,73] as planners and implementers of economic and social policies. From the time the proposal for the wind farm was put before the Marsabit County Council (MCC), the LTWP investors had a free hand in matters related to land acquisition, making concession agreements and defining which local community groups qualify as "indigenous" and hence qualify for compensation for the wind power project development in the area.

The initial application for the lease of 100 acres of land by the LTWP from the Marsabit County Council was made on 20 November 2006. According to the Trust Lands Act cap 288, which was the applicable law for community land at the time, the MCC was required, upon receipt of such requests, to notify the Divisional Land Board (DLB) or in the case of its absence establish one to meet to consider the proposal, notify people in the concerned area, hear and record submissions from people who were present and submit written recommendations to the council [74]. By the LTWP's own admission, public consultations began a year after their initial application, but no records exist to that effect. More importantly, the said public consultations were neither representative of the Loiyangalani residents, nor did they involve full disclosure of the project, as explained by the attorney representing the communities' interest in the case:

"The only meeting held was attended by town dwellers and fishermen from one area (Loiyangalani) and did not involve the pastoralist community and the project therefore lacks public participation in its establishment. This meeting neither declared the size of land required for the project, nor did it inform the community of the loss of access to their land, but instead concentrated on the benefits to the community." [70]

On 13 August 2017, the LTWP's application for a 99-year lease of 150,000 acres of community land was approved. Without a clear response from the MCC on whether the DLB existed at the time of the application and given the scant details of the community consultation carried out for the LTWP's application, the communities denied any knowledge of the project or its application for their land prior to site preparation activities in 2007. In its defense, the LTWP submitted that the instituters of the suit "were not inhabitants of the affected area ( ... ) that one can only have ancestral rights if one was a descendant of known ancestors and a denizen of the apposite area ( ... ) therefore cannot be representing the project affected communities [8]." Consequently, the LTWP invested heavily in legal representation to delegitimise indigenous communities' claims by promoting and actively pursuing an ideologically-charged discourse informed largely by the colonial conception of the communal utilisation of land. In responding to the claims of illegal annexation of community land during the inception of the project in 2007 and subsequent alleged unprocedural acquisition of an additional 110,000 acres of community land, LTWP used frontier imaginations of "idle, unoccupied" land to explain the rationale of dispossession. The following quote from LTWP lawyers at ELC proceedings illustrates this point:

"Mr. Nyaoga (LTPW lawyer) told the court that the land in question in this matter is currently uninhabited. He told the court that this would be made clear if the court visited the suit land. Mr. Nyaoga opined that the community was not merely being left on the roadside (Civil Suit 163 of 2014 (Formerly Nairobi ELC 1330 of 2014) | Kenya Law Reports 2018. Page 14 of 28. http://kenyalaw.org/caselaw/cases/view/130470. Accesses on 29 June 2018)."

More importantly, alongside the lack of community consultation, the LTWP's compensation entitlement of the affected communities' agenda reveals inconsistencies with the *Trust Lands Act*, drawing attention to instances of 'bribery' involving local political leaders [63]. Attributed largely to the fact that LTWP project designated the land as "empty" and because of its narrow definition of indigenous communities in the area, no compensation was paid. Instead of the mandatory compensation entitlements where community land is acquired, as stipulated in the *Trust Lands Act*, LTWP employed a predominantly Corporate Social Responsibility (CSR) focus (explored in the next section) that highlights the advantages of investments in community projects. The systematic presentation of CSR as "development gains" for the local community highlights the ways in which transformative CSR investments in infrastructure that support local livelihoods are linked to the LTWP project's wind power development in the area. CSR investments projects are chosen and refined through a "deliberative" process of consultation providing community "validation", perhaps the reason, despite the largely popular opposition to the project, that they remain popular.

Another feature of the LTWP project's development discourse is the conception of "local communities" as a simple agglomeration of "ethnicities" which, according to Ferguson, reduces the complexities relating to resource access and use to the "level of individual 'values', 'attitudes' and 'motivations' [4]." This conception was evident in the planning of the wind power project and in drafting an Indigenous Peoples Policy Framework in 2011, in which LTWP notes:

"It is clear that while most of these tribes are considered marginalised at international, regional and national levels, they have the same chance under this project to voice their concerns if their rights, interest, needs, livelihood, culture or desires are affected (Quoted in Danwatch report "A people in the way of progress")." [70]

In this respect, LTWP's conception of a picture of local communities as merely "ethnically diverse" takes no account of the competitive environments that characterise resource access and entitlement in a CPR system. In ethnically diverse settings, such as the LTWP project site, resource

access and entitlement can not only accentuate contestation and conflict between ethnic groups but also consequent market integration can lead to rising inequality and declining resource conditions for local livelihoods [75]. Apart from depoliticising natural resource politics and repositioning local ethnic groups as driven by nothing more than individual motivations, these representations assume that LTWP's "same chance under this project to voice their concerns" strategy will be free from social and political contestations inherent in this context.

#### (c) CSR—greenwashing dispossession

Claims of dispossession and contestation have affected the relationship between the company and the local pastoralist communities and polarised the local political leaders along the lines of ethnicity and whether a specific community supports the project. The combined effect of the representation of local communities and heightened ethnic tensions over the ownership of land in Sarima generated an ingrained antipathy towards the company, which the local communities view as a threat, occasioning economic and social challenges and hardships and "exposing their lives to vagaries of nature (Ibid Page 17–18 of 23)." The LTWP, in return, has sought to counter this antipathy through CSR investments mainly carried out through the Winds of Change (WOC) Foundation to achieve local level legitimacy and challenge the image of its label as a "land grabbing" corporate body. In effect, the CSR strategies seek a "social license to operate (SLO)" to "gain the approval and support of local communities" through "responsible" or "sustainable" management of the adverse social and environmental impacts of corporate activities [76,77] and to achieve legitimacy in their operations [78]. Hence, by establishing and funding the activities of WOC, the LTWP has a stated aim of improving the livelihoods of the communities in the project area (Winds of Change is expected to be funded for a period of 20 years (the operational life of the project) with a contribution of Euro 10 Million. WOC is expected to confine its operations to the Laisamis constituency in Marsabit County). According to the LTWP, the WOC activities will be confined to education and vocational training support, supporting health education and health infrastructure, and provisions of water to improve livelihoods with a shift that emphasises midterm livelihood activities. According to WOC website:

"WOC works in partnership with the county government, local leaders, NGOs, CBOs and government departments in implementing negotiated activities to ensure optimal stakeholder engagement, participation and ownership. WOC aims to catalyse positive sustainable development to enhance livelihoods in the areas surrounding the wind farm (WOC website https://ltwp.co.ke/winds-of-change/)."

For the LTWP, becoming "trusted partners" of local communities is important for two reasons. First, it is instrumental in seeking the recognition of, and, by extension, the legitimacy to operate in the area, through funding "negotiated activities". Basic health and education infrastructure has lagged in the region, and it has had one of the lowest human development indicators in the country [79]. The region, which has experienced increasing populations and associated competition for resources, has also been experiencing increasing droughts that have had constraining effects on the livelihood options and have generated the massive socioeconomic differentiation of households [51,80], significantly alternative livelihood trades, petty businesses and wage labor in the informal sector [81]. In the context of unpredictable weather, investments in education, healthcare and local livelihoods are thought to increase local communities' resilience to drought. To reduce livelihood vulnerabilities and improve the provisioning of basic services in the regions on the margins, development interventions work with and through existing policies that support the provisioning of basic services of education, health and entrepreneurialism [51]. The LTWP's CSR strategy therefore forms the "central core of a layered approach in which each layer relates to specific stakeholders and their associated concerns" [76], which is meant to legitimise the activities of the company in the minds of local communities. According to WOC,

"It is anticipated that the planned social investment activities will enable LTWP and its project partners to become trusted partners in development with the local community around the wind farm and the larger Laisamis constituency (WOC (2018). Community projects. https://ltwp.co.ke/winds-of-change/)."

This strategy has proved to be very effective, with WOC investing in over 30 projects in the Laisamis Constituency that span the education, health and water sectors. A health center nurse explained the WOC investment in health infrastructure and the implications this has had on the availability of essential drugs and health indictors in the area:

"We never had lights (in the health facility). I was using a spotlight to do deliveries (at night). Now we have a working solar powered lighting system and a delivery bed. We can now store the drugs here safely in this facility. The health indicators are going up (Interview\_024\_Health Centre Nurse, Video "Insights in to the social footprints of the Lake Turkana Wind Power Project" https://ltwp.co.ke/winds-of-change/)."

The appropriation of CSR to obtain SLO by the LTWP in the context of a contestation of its activities can be understood as examples of a "green washing" dispossession. However, as with Loperena's account of similar practices in conservation [82], CSR does not stop the exploitative processes of appropriation by the state and corporate investors set in motion by development visions of the state focused on appropriating natural resources premised on low-cost green energy development. Apart from the fact that CSR activities risk presenting certain business values as non-negotiable universal values and hence overriding the role of business in poverty reduction in the developing world [83], CSR activities, in a context where the state has failed to provide the necessary social amenities, as Ferguson argues, may reinforce and expand "the exercise of bureaucratic state power, which takes poverty as its entry point" in launching an intervention that has adverse effects on local communities [4]. The LTWP's CSR strategies mask the material practices of dispossessions of pastoral communities of their CPRs through CSR investments. The company's CSR investment strategy suggests that there may be important commonalities with past experiences where CSR are not adequately aligned with those of the local livelihoods [83] and do little to redress the pervasive resource conflicts as a result of the degradation and shrinkage of the natural resource base associated with LSLAs for incompatible development projects [84]. These pastoral CPRs are important because they harbor important resources for local people's livelihoods, enabling resilience against complex ecological contexts.

#### **7. The LTWP's Anti-Politics Machine and Comparative Observations**

The case of the LTWP has played an important part in the discursive process of endorsing and legitimising the development discourse through the reproduction and circulation of their own version of pastoralism representation vis-à-vis wind energy in Northern Kenya. Thus, this section provides a detailed commentary on the theoretical premises of the development representation, together with their institutional rationales through Ferguson's anti-politics machine framework: (1) the context in which anti-politics of dispossession of pastoral commons are constructed, (2) how the anti-politics machine is developed and (3) the key elements of the anti-politics machine, focusing on the articulation of the notions of "development" and "green energy" by the LTWP and the specific ways they are constructed around pastoral land use.

First, the context under which the LTWP has supported and promoted an anti-politics machine involves a complex array of historical and contemporary reproductions of pastoral land regions as "backward" and "emerging" [85]. Historically, pastoral lands have been seen by the colonial states as sites of famine and impoverishment, undermining political stability through forms of rebellion and insurrection [51]. The premise of pastoralism development discourse as an exercise in dealing with "backward, primitive and war-like" nomads through visions and plans entailing "modernity and progress, security and stabilisation" goes back to the colonial project in east Africa that sought to appropriate land for the colonial settlers. The position of the states in postindependence east

Africa changed very little in its policy discourse about pastoral land use. Apart from being viewed as "unproductive", states have viewed pastoral areas as a threat to the political, security and commercial interests of leading industrialised countries, in effect creating a precarious politics and, with it, top-down development interventions that often take a security dimension [51]. These trends, driven by neoliberal development policies encouraged by the World Bank [86] and bilateral donors, promoted interventions that focus on alternative land uses to pastoralism, including the promotion of private rights in land that not only led to the loss of communal land rights but also encouraged sedentarisation of pastoral populations, which has been associated with impoverishment, resource competition and population pressure [87,88]. Thus, even with evidence of massive failures of development interventions based on the incompatibility between modernised interventions and transhumant herders [89], the LTWP project utilises the state policy discourse that emphasises "opening up" marginal areas through large-scale investments in infrastructure and energy projects. Hence, the failure of the LTWP and local governance institutions to adhere to the guidance on stakeholder engagement and FPIC and the subsequent depoliticisation of wind power development in Northern Kenya is linked to a historical development discourse about the viability of pastoralism and communal tenure in the debate on land redistribution.

Second, the way in which the anti-politics machine has been developed within the LTWP's development discourse involves alliance building with state agencies and local elites that is meant to downplay the political character of land redistribution. The processes of land acquisition for the wind power project have been generated less through community consultation established in the law and more so through smaller and select groups of local and national institutions and elite networks. This was described in detail in the defense statements of the LTWP in the ELC:

"He told the court that the first defendant (LTWP project) approached the then County Council of Marsabit with its intention to embark on a wind energy project in 2005. He said that from the year 2005, continuous consultation between the defunct County Council of Marsabit and the local community had taken place. He also told the court that the defunct County Council of Marsabit and the National Government had been in consultations for a period of 9 years." (Records of the ELC proceedings on 8 March 2016 [8])

The key point here is that the processes of land redistribution are partly in continuity with previous interactions between the state and pastoral communities [90], where the state has perpetuated dispossession without regard for legal procedures. In this sense, the LTWP's anti-politics machine relies on being aware of and working within a broader state development discourse on "pastoral life-worlds, politics and resource management [85]."

The core elements of the LTWP's anti-politics machine are representations related to "green energy" development and pastoral land tenure. This discourse serves the dual purpose of representing the transformative impacts of the wind power project on pastoral land on one hand and, on the other, a representation of pastoral land use as "idle" and that needs to be "opened up", a discourse that tends to propagate a depoliticised interpretation of pastoral land tenure and resource governance, which are products of colonial antagonisms towards communal land rights. The LTWP has provided several representations that illustrate the importance of "affordable green energy" for the "development" of the region, creating an interdiscursive process where dispossession of "wasteful" pastoralists and "idle" community land is justified for the greater good of "development". This strategy shifts the focus of grassroots resistance from corporate actors to public institutions, which is presented as a neutral tool for planning and implementing economic policy and is incapable of working against the interests of the people. In constructing its anti-politics machine, the LTWP's strategy of low-cost green energy development is discursively constructed in ways that emphasise the backwardness of the region and the investments required for its transformation, without any mention of the project's social, economic and environmental impacts on the local communities. Once constructed, the anti-politics machine "shapes not only the formation of reports and documents, but the construction of organisations, institutions

and programs [4]." For instance, the ELC, in referring the case for hearing and determination by a bench of judges, elaborated the court's position:

"A question arises as to whether the alienation and annihilation of the community's ancestral land to the tune of 150,000 acres amounts to an irreparable loss and an unquantifiable inter-generational loss (......) that the project is the single largest private investment in Kenya's history, and that through the Power Purchase Agreement, public funds have been and continue to be utilised (......) The fact that the project is massive appears not to be in dispute. (......) I am inclined to find that the manner in which public funds (which is the tax payers' money) have been used and continue to be used in a project of such great magnitude is an issue of general public importance and of great public interest." [57]

Thus, a discourse is built around government policy on the importance of efficient utilisation of public finances that would support national development. The LTWP's anti-politics machine builds on these efficiency motives and builds a cumulative discourse about how 'low-cost green energy' will benefit national development. This reiterates Ferguson's insightful observation that it is the "promise of real input that makes the "development" form of engagement such a tempting one for many intellectuals [4]." The anti-politics machine not only represents the LTWP as a dutiful private sector player committed to delivering low-cost green energy but is also charged with the task of empowering the region's economically marginalised poor.

Complicating matters is the fact that community opposition to the LTWP reflected ethnicity-centered relations in the area. The litigants in the ELC case against the LTWP were mainly drawn from Rendille and Samburu pastoral groups that claimed the project was not only going to affect their livelihoods through reduced access to dry season pastures but would also reduce access to ancient ancestral sites for cultural rituals (Important cultural ceremonies of the Rendille and the Samburu include initiation rituals, *galgulame*—a gathering of all Rendille clans once in 14 years, a year after the circumcision ceremony. For a detailed description, see Schlee (1989) [91]). The LTWP's articulation of "community" in the area was drawn from a narrow definition as follows:

"Sarima village is the only village located on the wind farm. The ESIA process established that the location of Sarima village on the C-77 road would pose health and safety risks to the community from increased road traffic during the project's construction phase." [92]

Under the company's own admission, the Turkana population in the area only settled at the project site in the mid-2000s—the same time the project was initiated—after experiencing heightened conflicts in Turkana County [92]. This trend has been manifesting in the pastoral areas of the Horn of Africa, where Goldsmith [93] observed that high rates of demographic growth and contested rights because of uneven capitalists' transitions in the region are leading to conflicts. Similar dynamics have been observed in Northern Kenya [94] and Southern Ethiopia [85], where economic incorporation is promoting new forms of criminality and the politicisation of kinship relations, respectively. This, according to Hagmann and Mulugeta, is a characteristic trend of the securitisation of development projects in pastoral areas, where a section of the local community or group or their local leaders is coopted to uphold security and state interests at the local level [85]. In the case of the LTWP, the illustration of Sarima as the "only village located in the wind farm" has not only excluded the larger pastoralist communities dependent on the region for livelihood and cultural reasons but also served to redefine ethnic communities competing for benefits from the project. According to a local NGO,

"The company (LTWP) seems to adopt a de facto policy of encouraging the Turkana to settle on a land that is not clearly theirs. First, they denied the Rendille and the Samburu who are the real owners of this land their rights for 'living far from Serima'. Then, they claim the same pastoral land tenure allowed the Turkana to own land that doesn't belong to them (even though the company acknowledges Turkana's settled here in the 2000s). The selective representations of pastoral land tenure points to just how dishonest this whole arrangement is." (Interview\_013\_Local NGO representative interview in Nairobi, 7 May 2017)

Likewise, in terms of CSR investments, the WOC newsletter has been an important avenue through which to disseminate the work carried out by the company that benefits the local community from the project through the commitment to "invest a portion of its operating revenues to WOC to improve the livelihoods of the communities in the project area" [95]. These types of striking selective representations of local communities by the LTWP have provoked differing reactions from the local communities, some of which have asked to be enjoined in the suit against the company as "interested parties" that can attest to the benefits of the investment to the local areas but also legitimise the company's discourse on pastoral land tenure. Through their attorney, the interested parties, who are the residents of Loiyangalani District, argued that:

"the proposed interested parties were the ones directly affected by the project. He said that they had been consulted in all matters concerning the challenged project and that the consultations started around 2005.... the plaintiffs/applicants were strangers. He said that for the first time since independence, the interested parties were seeing the fruits of independence through the challenged project.... (they implored the court) to maintain the status quo (Civil Suit 163 of 2014 (Formerly Nairobi ELC 1330 of 2014) | Kenya Law Reports 2018: Page 20 of 23. http://kenyalaw.org/caselaw/cases/view/130470. Accesses on 29 June 2018)."

Under this discourse, the frequent selective representation of pastoral commons by the LTWP has a reiterative effect that is important in the establishment and perpetuation of the anti-politics machine. This discourse can also be read beyond the company's CSR activities and understood in terms of the representation of ethnic ascriptions and ethnicity as "economic assets in a development process" [96] within ethnically mixed settings.

In sum, the LTWP is the key actor in the wind power development in Northern Kenya, in terms of constructing, sustaining and reproducing the anti-politics machine that this paper has identified. This contribution has identified the reproduction of historical misconceptions about pastoral land tenure "progressively governmentalised" [4,97] by the bureaucratic state institutions, but also found the specific knots of depoliticisation of development interventions conceived, implanted and legitimised by the LTWP. Its reproduction of the Kenyan state's discourse on key discursive elements, such as 'low-cost green energy' and representations of pastural land tenure, not only guarantees continuity with previous interactions between the state and pastoral communities largely experienced in the east African region [85,90] but also shapes the larger mechanisms through which such misconceptions are produced and reproduced in development interventions. In this way, the LTWP's anti-politics machine provides a medium for the expansion and depoliticisation of state development visions. *The Vision 2030* and its flagship project—the LTWP project—have provided an apparently technical and apolitical entry point for an intervention that largely dispossesses communities of their land.

The analyses in this paper provide insights into the extent to which the processes of depoliticised development discourse identified in this single Kenyan case operate in other pastoral commons development contexts. Certainly, there are a lot of context-specific factors that are particular to the Kenyan case presented in this paper that limit the generalisability of this case study to other similar contexts. For instance, unlike the exclusion of communities in land acquisition in the Kenyan case, in Ghana customary authorities were proactively engaged by a private company in the process of land acquisition and benefit sharing in a biofuel plantation project without any government intermediaries [98]. Other experiences across similar LSLAs in Kenya and similar African contexts indicate that the pattern of grassroots mobilisation is not always necessarily one of "expose and oppose" as it was with the LTWP project case, but of attempting to ensure greater inclusion, incorporating the improvement of the terms of incorporation and recognition of claims over territory. For instance, the case of overt protest by the local Turkana community against the oil exploration company Tullow Oil Kenya sought more jobs for the local community members and a bigger share of the oil revenue to the region [5]. However, there are similarity factors that support the view that some degree of generalisability may be possible from the case of LTWP project in Kenya.

In post-colonial Africa, the persistence and centrality of mobility and communal land ownership were associated with "the tragedy of commons" [21] which governments in Africa used as justification for taking away rights and responsibilities related to resources out of the hands of local communities [99]. Further, communal land has been pointed to by state development policies in developing countries as evidence of "marginal", "empty" and "available" for alternative development [42,100,101]. These categorisations of pastoral land use and common property rights have become key operational mechanisms through which land-use changes are facilitated [102]. Hence, many development visions for the "development" of pastoral commons remain remarkably uniform and standardised, as illustrated by cases from Africa pastoral commons, both generally [2,39,64,100] and specifically in among pastoral commons in Ethiopia [6,103–105], and west Africa nations [106]. These examples are illustrative of the fact that the Kenyan case shows similarities to other practice associated with depoliticised representation of pastoral commons in very different contexts and therefore does not seem to be an exception.

Another aspect which could influence the generalisability of the Kenyan case is ideological underpinning of transformative development policies which is "premised on a classic conception of modernisation, which presumes a steady decline in the share of agriculture in the national income and in the composition of the labour force" where pastoral communities "constitute a vast reservoir of backwardness and a structural impediment to economic modernization [6]." The process of "development by dispossession" shows continuities with historical processes of dispossession across the world [107] with the vulnerability of lands to dispossession being highly correlated with the extent to which local and international legal systems recognise the multiple values that society attaches to land [42]. The Kenyan legal situation of communal tenure (especially prior to the 2010 constitution) is similar to that of many other countries where the proprietary character of the commons has been denied by both colonial and post-colonial administrations [108] and classified as "wastelands" and legally fully disposable by the state at its will [2]. These trends in the historical dispossession of pastoral commons and the role of weak legal recognition show strong resemblance across pastoral populations beyond the Kenyan case.

Finally, CSR activities, the central focus of the corporate anti-politics machine, constitute the discursive logic of assuming away and rendering unproblematic the depoliticisation of development interventions. This strategy of "greenwashing dispossession" [109] and "engineering consent" [110] is not unique either to the Kenyan case or commons dispossession; similar experiences have been observed in African contexts and globally in different sectors of LSLAs, such as agriculture, mining, energy and infrastructure [111]. CSR investments are thus an inevitable characteristic of LSLAs deployed to make development interventions politically feasible in the face of determined political mobilisation from affected communities.

#### **8. Conclusions**

This study has unveiled the anti-politics machine in the relationship between development visions and accompanying LSLA programs and the dispossession of pastoral commons in the current LSLA in Kenya. The analysis of the LTWP revealed that there is more to the recent discourse on the development of wind power in Northern Kenya than the imperative of generating more energy at lower cost to meet the increased demand in the realisation of *Kenya Vision 2030.* Drawing on Ferguson's anti-politics machine, the paper has shown that development visions of the hitherto marginalised Northern Kenya and their focus on large-scale infrastructure and energy programs are contingent upon the construction of ASAL livelihoods as unviable—a dominant colonial and postcolonial discourse that legitimises alternative large-scale investments as the ideal. Accompanying practices, such as placing the responsibility for the planning and operation of LSLA programs into the hands of private sector actors that attempt to absolve state and local institutions from their responsibility and accountability projects, legitimises the depoliticisation of development programs to secure the extraction of value from natural resources by the private sector. LSLAs are constructed as the necessary interventions required to bring

the isolated ASALs to integration with the national economy, excluding local pastoral livelihoods from being recognised as viable investment options to economic development. This representation, through discourses, technical recommendations, institutional apparatuses and apolitical interventions, forms the anti-politics machine that depoliticises important realities that characterise CPR use and governance and makes them amenable to solutions that may have no effect on their sustainable development but instead have undesirable results. As Agrawal argues,

"Without attention to the politics that generates underdevelopment and environmental degradation as universal problems, it may be impossible to address poverty, underdevelopment, and environmental degradation effectively." [112]

This anti-politics machine has the undesired effect of prioritising natural resource appropriation for national development goals over local priorities for them. In the LTWP case, the state and its network of corporate actors and local elites uncritically endorse the acquisition of community land for wind power development through a process that plays down the significance of the inherent complexities of CPR use and governance and their contestations related to disproportionate access and control by local communities. This is achieved through the anti-politics machinery that enables the implicit reproduction of historical misconceptions about pastoralism and communal tenure to legitimise the alleged irregular acquisition of community land.

To build on the concept of an anti-politics machine and its role in dispossession of commons, putting some of our findings in perspective is necessary. The case of the LTWP project is emblematic of the depoliticisation of new development visions involving the opening of frontier borderlands as new sites for the investments needed for growth of the national economy. Although these development visions involve strategies promulgated by the state, they involve a broad range of actors and a combination of public and private finance and governance [64] through public–private partnerships [113]. The contingent nature of these development visions as "arenas of action" relies on what Scott refers to as "state simplification" processes involving the reorganisation of natural resources for easy manipulation by the state and the uncritical optimism in the possibilities of planning through which state development policies were facilitated. This means that the private sectors actors with the vital role of delivering these development visions have a high degree of political power within these arenas of action and may, and often do, engage in a depoliticised partnership with the state. In this context, unveiling the anti-politics machine strategies employed in development visions involving public–private partnerships is key to understanding how private capital is involved in the distribution, multiplication and intensification of state power [4]. This is particularly the case for renewable energy systems built to further capital accumulation [114], where private capital with the technical solutions and capital drives the agenda for the implementation of environmentally friendly technologies for energy production.

Further, the paper has shown that common assumptions in development discourses promoting LSLAs involving public–private partnerships promote a depoliticised representation of local communities and oversimplification of competing interests and power structures at multiple levels. The depoliticised discourse, as Ferguson argues, "provides the technical point of entry" [4] for an intervention serving the political interests of corporate actors. However, the LTWP project's anti-politics machine is certainly not the source of the depoliticising discourse and strategy but rather, as Ferguson argues, one among the links in the mechanism that produces and articulates it [4]. Even more importantly, the logic of the LTWP project's anti-politics machine draws from the (mis)representations inherent in the state's own development discourse that have characterised ASALs development since colonial times. As a result, these misrepresentations often bring to the fore state-centric notions of the understanding of viability of marginal lands, such as the ASALs "powerfully informed by ideas about efficiency derived from neo-classical economics, and is rooted in the dominance of a particular type of commercial farming" [7] which lays the justification for alternative investments in land which serves the goal of "reinforcing and expanding the exercise of bureaucratic state power [4]." In the case study presented in this paper, the dominant storyline by the TP regarding the role of the wind

project in dispossessing communities through irregular acquisition of their land is ceding ground to one, advanced by the LTWP project and its network of government institutions, that transcends the logic of dispossession—generating low-cost green energy and the contribution of the project to the regions development through infrastructure that will connect the hitherto marginalised region to the rest of Kenya. Consequently, legal protection and judicial support sought by the communities affected often "veers away from questioning the fundamental roots of land-grabbing...while engaging in the problematic notion of win-win scenarios [113]."

The case of the LTWP illustrates some of the tensions between land redistribution and depoliticising development discourse about ambitious development interventions that have been a characteristic occurrence in Kenya's ASALs. However, the LTWP is not the only case, and indeed, there are several examples of comparable cases where indigenous peoples and local communities were dispossessed of their communal lands without industry guidance on stakeholder engagement and FPIC [47,103]. Beyond the likely dispossession of commons, depoliticisation has influenced debates and outcomes on compensations to communities affected by land redistribution apparatuses and ethnic tensions. The depoliticisation of land redistribution for private sector-led interventions carried out under the "progressivist and triumphal banners of development" [115] has already had far-reaching effects on pastoral commons; it leads to the development of exploitative relationships between corporations and communities through the concomitant restructuring of CPR governance and to associated losses of important pasture and migration resources through the erosion of CPRs [116]. This challenges the resilient use of the pastoral rangelands, decreasing the potential for innovatory husbandry [115] and effectively constituting "resilience grabbing". The broad distribution of CPRs and the existence of customary rules that guarantee use rights to communities mediated through an inclusive process of management has given pastoral commons a unique resilience against unpredictability and spatial and temporal distributions across the pastoral rangelands. Land investments that extinguish use and access to critical dry season resources that enable transhumance and offset drought risk [117] and the erosion of an associated institutional design that ensures excludability and addresses the problems of subtractability [40] by extension actually generate greater vulnerability and poverty.

This paper has shown how seemingly simple technical solutions in "development visions" and accompanying public–private partnerships in Kenya employ a depoliticised representation of local realities in the ASALs, aimed at reinforcing historical ghettoisation of pastoralism and communal land tenure. However, considering the specificity of the Kenyan case, important questions not addressed in this paper include the following: Is the corporate anti-politics machine dynamic and relational or static? How is re-politicisation negotiated and with what impacts? This work indicates the need for further research on how the anti-politics machine may be differently generated and negotiated in the context of development visions.

**Funding:** This research was funded by Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung grant number P1BEP1\_175227.

**Conflicts of Interest:** The author declare no conflict of interest.

#### **References**

© 2019 by the author. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

### *Article*


**6. Possibilities of the presbycusis prophylaxis**

At the moment, the presbycusis is irreversible; therefore, the prevention of age-related hearing loss must be paid attention to. First of all, it is necessary to educate the population about harmful factors affecting hearing throughout the life, such as ototoxic drugs, noise exposure, vibrations, and others. Audiological care in case of SNHL should be aimed at the enhancement of nonspecific resistance of excitable structures of the auditory analyzer to general pathological damaging factors—tissue hypoxia, oxidative stress, and extinction of the action of endogenous neurotrophic factors [30, 83].

Due to the fact that with age-related hearing loss, it is impossible to obtain gains in tonal hearing, and special emphasis is done on means of improving the auditory attention (the functions of the central auditory pathways), which allows to compensate for the lack of auditory acuity and enhance the efficacy of the hearing aid. If CAPD causing poor speech intelligibility is detected, the auditory training is appropriate.


3.2.6. Inhibitors Screening, ADP-ribosylation of PARP-3

The reactions were performed in the presence of 2 mM MgCl2 as a cofactor in HDB buffer. The reaction mixtures (final volume 10 µL) contained 0.1 µM DNA substrate, 0.5 µM PARP-3, 5 µM [ 32P]-NAD+ and 0, 35 or 95 µM NAD+ in the absence or presence of various concentrations of the inhibitor, which are indicated on the figures. The reaction mixtures were incubated for 20 min at 37 ◦C then terminated by adding the 5x gel loading Laemmli sample buffer and heating at 85 ◦C for 10 min. The mixtures were resolved on a 10% SDS-PAG as described in [84] The gels were dried and subjected to autoradiography and/or phosphorimaging for quantitation using the Typhoon imaging system from GE Healthcare Life Sciences. Quantitative processing was carried out using OriginPro7.5, Microcal Software, USA. In all experiments, the points on the experimental curves represent the average of a minimum three independent experiments. Standard deviation did not exceed 10%.