Living cells are constantly exposed to mechanical stimuli arising from the surrounding extracellular matrix (ECM) or from neighboring cells. The intracellular molecular processes through which such physical cues are transformed into a biological response are collectively dubbed as mechanotransduction and are of fundamental importance to help the cell timely adapt to the continuous dynamic modifications of the microenvironment. Local changes in ECM composition and mechanics are driven by a feed forward interplay between the cell and the matrix itself, with the first depositing ECM proteins that in turn will impact on the surrounding cells. As such, these changes occur regularly during tissue development and are a hallmark of the pathologies of aging. Only lately, though, the importance of mechanical cues in controlling cell function  has been acknowledged. Here we provide a critical review of the recent insights into the molecular basis of cellular mechanotransduction, by analyzing how mechanical stimuli get transformed into a given biological response through the activation of a peculiar genetic program. Specifically, by recapitulating the processes involved in the interpretation of ECM remodeling by Focal Adhesions at cell-matrix interphase, we revise the role of cytoskeleton tension as the second messenger of the mechanotransduction process and the action of mechano-responsive shuttling proteins converging on stage and cell-specific transcription factors. Finally, we give few paradigmatic examples highlighting the emerging role of malfunctions in cell mechanosensing apparatus in the onset and progression of pathologies. ChangesThe best example of how pathology-driven changes in ECM mechanics and architecture impact on tissue and organ function is cardiac remodeling. Following a cardiovascular event , cardiac matrix is degraded and substituted by a scar, having a different nanostructure and compliance . The strSimilar effects of altered ECM mechanics on the function of different cell types have been recently demonstrated .The concept that cells can interpret and respond to mechanical cues is not exactly new to the scientific community. Nonetheless, only lately, the elucidation of the molecular mechanisms by which the cell perceives and transforms the mechanics of the ECM has become the subject of intense investigation and a number of intracellular molecules has been identified that can react to mechanical stimulation and - in turn – modify cell function.So far, the definition of cellular mechanosensor has been applied to a number of molecules – mainly proteins – displaying a status change in response to mechanical stimulation. The nature and the degree of the change imposed by mechanical cues can vary significantly and post-translational modifications , intraceCells perceive mechanical stimuli through diverse mechanosensitive molecules at the cell membrane including integrins, stretch-activated ion channels, G protein coupled-receptors, growth factor receptors, activating different mechanotransduction pathways .In the present review, we focus on the cellular mechanical response through ECM-integrin-cytoskeleton-nucleus axis and critically discuss the molecular basis of focal adhesion cell mechanosensing. We highlight how different intracellular molecules respond to mechanical loading and transfer the information from the very site of cell-ECM interaction – the membrane – to the nucleus, where the mechanosensitive genes are eventually activated.The primary site of force transmission to the cell is the cellular membrane, where the direct contact with the extracellular matrix (ECM) occurs.Cells in contact with a stiff surface typically develop discrete multiprotein complexes under the membrane named focal adhesions (FAs), which are the main hub of cell-ECM interaction.Focal adhesions mechanosensing activity consists in perceiving and transferring the mechanical cues arising from the extracellular milieu to the cellular cytoskeleton. To do so, they are built as complex structures, that can be divided in a transmembrane and in an intracellular layer. The intracellular layer of FAs is composed by scaffolding, docking, and signaling proteins that collectively serve as interface between the transmembrane components directly contacting the ECM (integrins) and the actin cytoskeleton. The molecular composition of FA core is extremely variable and sensitive to ECM composition and mechanics, as perceived by integrin binding. In fact, different degrees of integrin clustering, as determined by the spacing and availability of ECM adhesion sites, affect the recruitment of FA proteins to the binding site . Among tFigure 1.Given the complexity of FA structure and the number and nature of the proteins involved, the modalities by which the FAs act as primary mechanosensor cannot be described collectively; thus, the response of few key FA components to mechanical loading will be herewith described. A representation of the mechanosensing machinery of the FAs is depicted in The amount of tension generated by a given FA is thought to directly correlate with its size and with the amount of structural, docking and functional proteins recruited to the site .Focal adhesions are dynamically built following the assembly of transmembrane proteins deputed to physically interact with components of ECM, namely fibronectin, vitronectin, collagens, laminins, and named integrins. Integrins are heterodimers composed of α- and β-subunits, whose assembly is guided by ECM composition and whose specificity is given in mammals by the combination of 24 α- and 9 β-subunits and by alternative splicing events. The combination of α- and β-subunits defines the affinity of the receptor for different ECM components and its cell type specificity.Integrin affinity for its ECM ligand can be regulated within the cell in a process called “inside-out signaling” or prompted by extracellular mechanical stimuli, inducing a high-affinity conformation change . FollowiExtracellular matrix composition drives precise integrin subsets expression which, being coupled with different signaling cascades, induces specific cellular responses .Apart from being responsive to changes in ECM biochemical composition and mechanics, cells also adjust their own mechanical state by altering cytoskeletal architecture, modulating cellular elasticity, or generating a concomitant contractile response to applied forces . The intFocal adhesion kinase is one of the first molecules recruited to developing FA in response to external mechanical stimuli. Its activation by autophosphorylation is considered the trigger to intracellular mechanotransduction, by activating downstream mechanotransducers within the cytoplasm . DownstrTalin is a 270 kDa protein composed of an N-terminal globular head, a flexible rod domain and C-terminal helices. While the helices are involved in protein dimerization , the heaThe most recognized effect of force loading to talin consists in its unfolding to expose cryptic hydrophobic binding sites to host vinculin head . In the Vinculin is one of the main components of FA inner core and its recruitment to the site requires talin activation by mechanical forces : vinculiAnother model describes a more complex activity: when recruited to FAs, vinculin couples cell area and traction force with differential contribution coming from the head and tail domains. In fact, vinculin transmits force inside-out by increasing ECM-bound integrin–talin complexes via the head domain, while the tail domain is needed to propagate force to the actin cytoskeleton .Paxillin is a 70 kDa phosphotyrosine-containing docking protein being mainly localized at the FA intracellular layer. Here the protein is traditionally believed to integrate mechanical cues arising from the ECM and biological signals propagated via the growth factor receptors. The protein contains different interacting domains  which confer paxillin high-affinity binding properties to bring together structural and signaling partners . The mecWhen extracellular tension is reduced, FA sites lose the ability to recruit paxillin and detach from the relaxed substrate. This event abrogates actin polymerization, resulting in slow actin recovery and increased incidence of stress fiber breaks . PaxilliZyxin is a 61 kDa phosphoprotein containing three C-terminal LIM domains and nuclear exclusion sequence (NES). The presence of such domains accounts for its localization to the FAs and for the interaction with a number of FA partners. Zyxin mechanosensing activity consists in its dynamic diffusion through different cell compartments: zyxin is released from FAs when cells are grown on a soft substrate or when the mechanical load is reduced by inhibiting the actomyosin interaction . StretchCas, recently proposed as a novel mechanosensor , microtubules (MTs) and intermediate filaments (IFs). Each of the components displays a highly organized structure contributing to intracellular organelle integrity and maintenance .Cytoskeleton contractility is ensured by F-actin sliding on the motor protein myosin II. F-actin and myosin II are held together by crosslinking proteins  in complex structures called vice versa , capping proteins, depolymerizing factors (ADF/cofilin), stabilizing proteins and crosslinkers contribute to control the architecture and the mechanical properties of the network .The main process by which actin cytoskeleton is stabilized by tensile force application consists in the inhibition of the actin severing activity of cofilin . When acROCK activation by RhoA also induces myosin II activation by direct phosphorylation of myosin regulatory light chain (MLC) mainly at the Ser-19 residue or by inhibition of MLC phosphatase (MLCP) . MLC phoFigure 2).Besides, ROCK directly participates in cytoskeletal stabilization: Rho/ROCK pathway activates the formin Diaphanous (mDia), which directly or through the Arp2/3 complex promotes F-actin polymerization   were described in mammals that display different localization, tissue expression and enzymatic properties. As described before, MyoIIA is responsible for generating traction force in order to stabilize FAs in a Rho/ROCK-dependent mechanism. Due to its fast turnover, MyoIIA allows rapid cytoskeleton remodeling. MyoIIB is, instead, an actin fiber stabilizer with no motor function; it is localized at the perinuclear actin cap and involved in maintenance of cell polarity .SFs are physically connected to the MT network . MTs, thIFs are highly flexible and more stable as compared to F-actin and MTs. Their dynamics and interaction with numerous signaling pathways are regulated by post-translational modifications .Table 1) several groups have identified kinases and transcription factors , showing that the Armadillo Repeat Region (ARM) is mechanically unstable and displays multiple alternative unfolding rounds .After the discovery of β-catenin shuttling ability, a number of other proteins have been shown to relocate to the nucleus following modifications in ECM composition and mechanics.Among the proteins sitting at the FAs in static conditions, and shown to detach from the membrane site and move to the nucleus following dynamic stretching, is zyxin . As descPaxillin is also credited of having a structural function at the adhesion sites while shuttling to cell nucleus in response to mechanical stress. This protein is predominantly localized to the FAs and its localization can be modified following different rounds of phosphorylation on tyrosine and serine residues by FAK in response to modifications in cell spreading and polarity . Its detA new class of shuttling proteins acting as mechanotransducers, by moving back and forth from the nucleus without being physically associated to FAs, has been recently described, which will be discussed in detail in the following section.Figure 3).Yes-associated protein (YAP) and WW Domain-Containing Transcription Regulator Protein 1 (WWTR1/TAZ) are transcriptional co-activators being the downstream effectors of Hippo pathway . In respThe definitive demonstration of YAP acting as a mechanosensitive protein was recently given through an elegant experiment performed by the group of Rocha-Cusachs: besides being translocated upon nuclear pore opening following the application of force on the cell, YAP shuttling to the nucleus was shown to depend on the intrinsical protein mechanical instability .YAP/TAZ paradigmatic ON/OFF switch-like behavior has been reported in a number of cell types and, if coupled to their acknowledged role in controlling organ shape and size during organogenesis, is the perfect example of how ECM composition and mechanics can impact organ function. YAP/TAZ persistence in the nucleus is regulated by the phosphorylation on specific Serine residues  operated by Hippo pathway upstream regulator LATS1/2 and can be released by dynamic modifications in substrate compliance or nanostructure . DistincSince the main annotation for YAP/TAZ transcription targets lies within the proliferation category, the activity of Hippo effectors in the nucleus has been historically associated with cell growth and tumor spreading , while oAlthough recent reports suggested mechanical signals influence the expression of mechanosensitive genes, the molecular processes by which mechanical forces are transmitted from the periphery to the nucleus of the cell, the largest and stiffest organelle in eukaryotic cells, are still largely unknown . In factThe existence of a connection between nucleus and cell membrane has been proven by experiments showing that the application of mechanical forces on integrin receptors at the cell membrane is followed shortly by nuclear structural reorganization and deformation in the direction of the pulling force  and by cThe nuclear-cytoskeletal coupling is crucial for force transmission to the nucleus and, consequently, for the biological response. Many studies have pointed at the nuclear envelope as a regulator of biochemical and physical connection between nucleus and cytoskeleton . Indeed,Figure 4). SUN proteins also interact with nuclear pore complexes (NPC) controlling their organization and distribution on the nuclear envelope  . SimilarLINC perturbation has been associated with actin cytoskeleton derangement , nuclearOn the internal side of the nucleus, SUN proteins link intimately to the nuclear lamina through the main stabilizer of the INM, the intermediate filament lamin A . This inIndeed, changes in lamin A protein levels have been detected when tissue rigidity is modified, with soft substrates inducing its phosphorylation and consequent dissociation from the nucleoskeleton . Lamin ASUN nucleoplasmic domain and lamin A are connected to nuclear chromatin, thus possibly affecting directly gene regulation . Lamin AEmerin has been recently described as a sensor of tension upon mechanical stimulation: in isolated nuclei exposed to mechanical stimulation, emerin undergoes phosphorylation by Src kinase, thus leading to lamin A accumulation at the nuclear envelope and nuclear stiffening . Cells lIn a similar fashion, LINC complex and nucleoskeleton components are involved in Wnt pathway regulation; emerin overexpression prevents β-catenin nuclear shuttling and its activity, whereas emerin-depleted cells show substantial accumulation of β-catenin in the nucleus .These results appear more interesting when corroborated by the evidence that lamin A and emerin associate with multiple factors involved in transcription regulation, chromatin organization and mRNA processing, thus implying that mechanical cues can impact on mRNA translation through LINC complex .The association of transcription regulators, LINC complex and nucleoskeleton components at the nuclear periphery has been linked to both the activation and the repression of transcription. Transcriptional activity has been correlated with chromatin rearrangement at the nuclear periphery, in particular to the interaction between euchromatin and NPCs. NPCs are indeed recognized as active transcription sites connected with both cytoskeleton and DNA .Another way by which INM proteins can modulate gene expression in a mechanosensitive fashion is represented by reducing the accessibility of chromatin to transcription regulators. Lamin A binds sites of transcriptionally silent heterochromatin at the INM, while transcriptionally active euchromatin is distributed at the center of the nucleus. According to this model, the nuclear periphery can serve as a resting site for transcription factors, sequestering them and preventing their interaction with target genes .C-Fos represents a well-described example of transcription factor being sequestered by lamin A/C at the periphery of the nucleus .The interaction of nucleoskeleton with transcription factors known to be directly activated by mechanical signals deserves more attention: Mega-karyoblastic leukemia 1 , member of myocardin family, is a mechanosensitive transcription factor which dissociates from G-actin in the cytoplasm upon mechanical stimulation and activates SRF in the nucleus. Alterations in nucleoskeleton organization affects MKL1 pathway as demonstrated by impaired MKL1 nuclear translocation in lamin A/C depleted cells .Also, NF-κB, which is mechanically induced to translocate in the nucleus, suffers defects in the nuclear-cytoskeletal coupling .The interpretation of mechanical cues by the cell is completed by the activation of a given genetic program which induces the cell to adapt to the new conditions.in vitro, while laminin seems to restrict their response  are grown onto surfaces displaying stiffness gradients. Consistent with the acknowledged ability of the cells to perceive different substrate stiffness, MSCs were shown to migrate toward the stiff area, in a mechanism dubbed durotaxis, which is dependent on cytoskeleton dynamics . Since sresponse , thus hiin vitro studies can lead to discrepancies and different interpretations of results. In order to have a clear overview, the experimental design must consider the stiffness range specific for each organ/tissue. Depending on the function in the body, softer tissues such as brain (1 kPa) and harder tissues such as bone (1 GPa) can be identified  as compared to microgratings (10 μm width) or unpatterned surfaces . The spaBy controlling nanostructured materials periodicity and spacing, as to match integrin size and spacing through nanodots of 8 nm, it was indeed possible to tune integrin clustering and cell adhesion. By increasing the spacing between the nanodots, integrin clustering was abolished and cell adhesion compromised . Similarin vitro cell confinement models have been proposed with the aim to reproduce cell constraints by controlling cell shape, area and spreading  with ECM at specialized Z-band sites named costameres. The discrete distribution of the costameres in correspondence of the intercalated disks and Z-bands appears as the most efficient way to ensure the transmission of the forces to the sarcomere; indeed, the derangement of the costameres is a common feature of dilated (DCM) and hypertrophic cardiomyopathy (HCM) .in vivo , limb-girdle muscular dystrophy], cardiopathies (dilated cardiomyopathy), metabolic diseases  and premature aging disorders  .Skeletal and cardiac muscular dystrophies are the laminopathies identified most frequently and include limb-girdle muscular dystrophy, autosomal dominant EDMD, and congenital muscular dystrophy .Animal experiments show that lamin A/C knock-out mice develop cardiac and skeletal muscular dystrophy and cells isolated from these mice show defects in the nuclear shape, the distribution of nuclear pore complexes and the mislocalization of nuclear envelope components, such as the inner nuclear membrane protein emerin . InteresThe molecular basis of the laminopathies are still debated. Due to the role of lamin A as a scaffolding protein of the nucleus, LMNA mutations can result in LINC organization impairment and, consequently, in defects in anchoring the nucleus to the cytoskeleton. Indeed, LMNA-mutated or knock-out cells show defective nuclear-cytoskeletal coupling, deranged nesprin-1 positioning and altered TAN line anchoring. They are thus more susceptible to mechanical stress . As expeTogether with its role as nuclear scaffolding protein, lamin A also functions as an anchor site for chromatin to the nuclear periphery. As such, it interacts with components of the transcription machinery. Therefore LMNA alterations can prompt chromatin derangement and changes in gene expression. Nuclear envelope defects, heterochromatin displacement from nuclear periphery and nuclear membrane fragility are common features in cells obtained from EDMD patients .Besides, laminopathies can result from defective lamin A processing . Lamin AThe consensus over the importance of mechanical signals in shaping cell and tissue function started building with the evidence that cell fate  and funcModifications in the compliance of ECM are typically associated with the onset and the progression of degenerative diseases  and are mechanotransduction”, a term which applies to all the molecular processes contributing to transform physical cues into a biological response , those transducing the information toward the nucleus (mechano-transducer) and those activating target mechanosensitive genes (mechano-actuator) would be beneficial in drawing the borders of this rather new discipline.Figure 5). In this context, the first step is universally recognized as the activation of integrins, which are bound to set the mechanosensing pitch at the nanoscale level  following the interpretation of the mechanical signals, shuttling mechanotransducers enter the nucleus and function as adaptors for cell- and stage-specific transcription factors; (2) mechanoresponsive transcription factors exist that are only activated or made available for transcription following alterations of ECM mechanics. Although the former appears more realistic in the light of the example of YAP/TAZ co-transcription activators (Additionally, very few studies addressed the modalities of activation of the mechanosensitive genes so far. Two hypotheses can be drawn, which are depicted in tivators , a systeAn important task for future research will be to elaborate integrated strategies aimed at unraveling the interactions among different mechanobiology pathways, which at the moment appear to be intertwined in a complex web .in vivo situation. This approach would eventually help building more reliable models of mechanosensing failure and identify pathological conditions due to the derangement of the mechanotransduction apparatus.Finally, a further challenge for the future will be represented by the need to scale up mechanobiology studies as to fit a 3D setting, in order to make them more predictive of the FM proposed the subject and conceived the general structure of the review. FM, AP, VV, and SP revised the existing literature and contributed to all the sections. GF revised the text and contributed the discussion and conclusion.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


The Indo-Gangetic aquifer is one of the world’s most important transboundary water resources, and the most heavily exploited aquifer in the world. To better understand the aquifer system, typologies have been characterized for the aquifer, which integrate existing datasets across the Indo-Gangetic catchment basin at a transboundary scale for the first time, and provide an alternative conceptualization of this aquifer system. Traditionally considered and mapped as a single homogenous aquifer of comparable aquifer properties and groundwater resource at a transboundary scale, the typologies illuminate significant spatial differences in recharge, permeability, storage, and groundwater chemistry across the aquifer system at this transboundary scale. These changes are shown to be systematic, concurrent with large-scale changes in sedimentology of the Pleistocene and Holocene alluvial aquifer, climate, and recent irrigation practices. Seven typologies of the aquifer are presented, each having a distinct set of challenges and opportunities for groundwater development and a different resilience to abstraction and climate change. The seven typologies are: (1) the piedmont margin, (2) the Upper Indus and Upper-Mid Ganges, (3) the Lower Ganges and Mid Brahmaputra, (4) the fluvially influenced deltaic area of the Bengal Basin, (5) the Middle Indus and Upper Ganges, (6) the Lower Indus, and (7) the marine-influenced deltaic areas. The Indo Gangetic Basin alluvial aquifer system (known as the IGB aquifer throughout the rest of this report) is one of the world’s most important transboundary freshwater bodies  was used to spatially interrogate the different types of information held within the database. Data were systematically reviewed to apply a confidence rank to the studies and the data contained. Such a review process was essential to ensure that the combined hydrogeological datasets and maps developed were based on data generated by comparably robust methods, scale and data quality, despite being of multiple third-party provenances. Key criteria used in assigning a confidence value were: (1) that the depth and aquifer unit for the study was well constrained; (2) the date was available for temporally dependent data ; (3) sufficient methodological description was available to assess the quality of the data; and (4) the methods used to generate the data were appropriate. Data which only partially satisfied these criteria were ranked to be of lower confidence. High-quality studies were used as the key inputs to develop and parameterize the new basin-wide maps of each aquifer parameter, and lower quality studies were used only in areas where there were few or no high-quality studies to use. The review process identified 80 key benchmark studies which provided the highest quality systematic regional data and these formed the key datasets for the typologies.Available regional and national geological maps, alongside the large number of published sedimentological studies, were used to develop a new geological map of the alluvium of the exploited IGB aquifer. This map provides higher-resolution information on the geological age and sedimentological characteristics of the alluvium of the effective aquifer across the IGB than previously available within transboundary geological maps of the aquifer system, which traditionally portray the aquifer alluvium as a single homogenous unit at this scale. Identifying this variability of the alluvium sedimentology at a transboundary scale provides a robust framework within which to consider changes in the hydrogeological properties of aquifer.Existing available national and regional geology maps were used to develop the initial line work of the map, and were used to identify and demarcate the spatial extent of differing ages of alluvium across the aquifer. This line work was then validated and parameterized with information on modes of alluvium deposition, likely grain-size and sediment composition from the large amount of detailed information contained in the many high-quality georeferenced sedimentological studies across the IGB aquifer at local-scale. By combining these data, nine discrete units of alluvium characterized by key systematic changes in stratigraphy, were identified and mapped across the aquifer.Copious hydrogeological data are available at local-scales across the IGB aquifer. Over 400 hydrogeological studies and datasets were georeferenced and quantitative data used to identify spatial patterns in transmissivity, aquifer anisotropy and to develop a map of specific yield.2. Systematic patterns in aquifer anisotropy were identified from existing data and published studies within the Upper and Lower Indus basin  and a review of local-scale hydrogeological studies and regional maps across the IGB aquifer. Values of transmissivity extracted from the studies were georeferenced and spatially interrogated in GIS. From this, broad areas of similar ranges in transmissivity were identified and delineated for both the Holocene and Pleistocene-aged alluvium. Due to the local-scale variability of the alluvium, areas of distinct transmissivity ranges were only delineated where transmissivity values varied consistently in comparison to an adjacent area by either >100% or <50%, and over a spatial area of ≥2,500 kmA different approach had to be taken to map specific yield (Sy) across the basin due to the local availability of specific yield data. Grain-size distribution data available from sedimentological studies for the upper 200 m of alluvium (the effective aquifer) across the basin were used as a proxy for Sy and mapped across the basin. Values of specific yield were attributed to the different grain size ranges based on studies on specific yield for grain sizes within Bangladesh  datasets for the years 1950 to 2012 ; groundwater chemistry ; and the dominant recharge process. Areas with similar properties were digitized to illustrate the extent of each typology.The report provides a résumé of the new basin-wide understanding of the IGB aquifer developed from the review, new transboundary maps developed of the key properties and the deduced typologies. The major characteristics of each the typology are described alongside the implications for groundwater governance.The Indo-Gangetic basin developed 15 million years ago in response to uplift of the Himalayan plateau with lithospheric loading and depression of the Indian continental plate , up to boulder-sized, close to the mountain margins of the IGB aquifer and within megafan deposits Day , to mediThe ratio of channel to inter-channel deposits within the Pleistocene and Holocene alluvium also changes across the IGB aquifer from proximal to distal areas, and is highly significant to the proportion of higher permeability sand deposits, to lower permeability silts and muds, which comprise the aquifer in different regions, and the anisotropy of the alluvial aquifer system.Within the upper and central parts of the IGB basin, rivers systems are incising and of lower lateral aggradation than further downstream Singh . As a reThe Pleistocene and Holocene alluvium have a distinct distribution within the IGB aquifer, which underpins many of the systematic changes in aquifer properties from proximal to distal areas in the aquifer system. In the upper and central parts of the IGB the effective aquifer is predominantly composed of Pleistocene-aged alluvium, as result of the strongly incising nature of rivers. Recent-Holocene aged alluvium is only deposited within narrow terraces (km wide) adjacent to active rivers , waterlogging and increased salinization can occur. Over-irrigation can also lead to degradation of groundwater quality , southern Nepal, the Sylhet trough in eastern Bangladesh, and within Holocene sediments along the course of the Ganges in northern India, and also within the Indus system Fig. . ThroughRecharge to the IGB aquifer is complex and often controlled by several factors in any one place including rainfall volumes and intensity, canal leakage, irrigation returns, river inundation and even abstraction. Three datasets relating to the main recharge processes of rainfall, rivers and canal leakage were developed to understand how groundwater recharge may systematically vary across the basin. Figure There is considerable evidence of high rates of groundwater recharge from seasonal rainfall within the aquifer. Where average annual rainfall is greater than 1,000 mm, such as along the northern, central, and eastern parts of the IGB basin, direct, rain-fed recharge generally dominates  the piedmont margin; (2) the Upper Indus and Upper-Mid Ganges; (3) the Lower Ganges and Mid Brahmaputra; (4) the fluvially influenced deltaic area of the Bengal basin; (5) the Middle Indus and Upper Ganges; (6) the Lower Indus; and (7) the Marine influenced deltaic areas. The distribution of the typologies is presented in Fig. The Piedmont is split into three sub-typologies: the main piedmont margin of the IGB aquifer along the edge of the Himalayas; the lower piedmont plain; and the inner valleys.The main piedmont margin forms a distinct narrow band of very high permeability coarse deposits of variable thickness that extend tens of km along the majority of the northern margin of the IGB aquifer in India and Pakistan at the edge of the Himalaya Middle Hills  is situated closer to the IGB plain and has distinct differences in hydrogeological properties from the upper piedmont typology Fig. . The typEnclosed valley settings in the lower piedmont, called ‘inner valleys’, are present in a few areas . The groundwater resource is of significantly lower potential in these small disconnected valleys, as a result of the limited spatial extent of the alluvial aquifer and the absence of a significant upper piedmont terrace Fig. . The per2 and is over 200 m thick. The typology is highly exploited with many shallow tube wells (<100 m), hand dug wells and a growing number of deeper tube wells  to 50–70 m/d (in sand dominated channel deposits). The typology has a high specific yield of 10–25% Chaudhri .At a local scale, aquifer properties can vary significantly over short (100s of m to km’s) distances and thicknesses. Low-permeability layers stratify the aquifer, but are rarely continuous over more than a few kilometres  and greater groundwater potential (25–40 L/s in tubewells 10–60 m deep) than the surrounding typology of the Upper IGB plain Fig. . The larThis typology forms the effective aquifer unit across a large part of the eastern part of the IGB aquifer, stretching across northeast India and north Bangladesh. The typology has notable differences in groundwater chemistry and recharge to that of typology 2 within the Upper Indus and Ganges IGB plain Fig. .Fig. 16SThe typology is formed of highly permeable, predominantly Holocene-aged fine-medium sand-sized alluvium Singh . HydraulGroundwater is widely used in the typology, particularly within Bangladesh. Across the typology, abstraction is widely focused to the upper shallow aquifer horizons 0–50 m bgl, as it is across the IGB, and shallow tubewells in the typology typically provide 20–70 L/s. Deeper tubewells, where employed, often provide higher yields, sometimes >100 L/s, and highlight the potential of the multi-layered aquifer.There are important differences in the groundwater chemistry and recharge in this part of the IGB aquifer, compared to the Upper Indus and Ganges typology (typology 2). Saline groundwater is not as widespread but elevated arsenic concentrations (often >50 μg/L) are common in localized areas throughout the typology  mean that there is limited accessible and safe fresh groundwater in the shallower horizons of aquifer despite high rain-fed recharge Fig. . As a reThe primary aquifer in the typology is formed from Holocene alluvium. Relative to upstream typologies, the Holocene alluvium is composed of a significantly higher proportion of silt and clay (50–70%), and the overall bulk hydraulic conductivity of the aquifer typology is lower, typically 5–35 m/d, and specific yield is <10%. The anisotropy ratio of the Holocene alluvial aquifer is high due to the greater prevalence of low-permeability units in the typology, which are continuous over tens of kilometres Jones . Mean hoElevated arsenic concentration in shallow groundwater is widespread and very high concentrations (>200 μg/L) are common , which receives significant rain-fed recharge, average annual rainfall in the lower Indus typology is low, <250 mm, and evapotranspiration is high IWASRI . GroundwGroundwater is used much less extensively in this part of the basin, and the typology is characterized by an area of groundwater-level rise in the IGB aquifer, in strong contrast to other parts of the basin, where abstraction, groundwater storage potential and recharge are higher.Both coastal margins of the IGB aquifer represent areas of relatively low fresh groundwater potential in the IGB aquifer system Fig. . SalinitFour minor typologies describe important aquifer variations along the margin the IGB plain aquifer. These include: the deeper groundwater on the Bengal Basin, the southern marginal alluvium in the Ganges basin, the western Indus basin piedmont, and the Sylhet trough in Bangladesh.Within the Bengal Basin, deeper groundwater (150–350 m mgl) is an important resource. Across the rest of the IGB alluvial aquifer system, groundwater is rarely used below 150 m mgl; hence, the restriction of much of this report to discussing the shallowest 200 m. However, due to the widespread contamination of shallow groundwater with arsenic and the evidence of low arsenic concentrations in groundwater > 150 m . The high specific yield of the IGB aquifer means that large volumes of groundwater are stored in the aquifer system—potentially, 20 times the annual flow of the river systems , and monitoring is essential for appropriate and timely policy frameworks to be established that enable a managed response. Continued exploration, testing and monitoring of shallow and deeper groundwater and their interaction with surface-water sources across the aquifer system are needed to enable timely management systems to be developed to identify and mitigate further degradation. The typologies, which highlight the current status of the IGB aquifer groundwater resource, as well as the major hydrogeological differences across the system, provide a robust framework to inform different capacity development and monitoring requirements within the different parts of the aquifer, to develop effective management strategies at local and regional levels.The hydrogeological typologies systemize groundwater knowledge of the Indo-Gangetic alluvial aquifer system, and assemble the information most relevant to understanding its resilience. The seven major typologies highlight significant spatial changes in recharge, permeability, storage and groundwater chemistry at a transboundary-scale across the IGB aquifer, and provide an alternative conceptualization of this aquifer system which is traditionally mapped as a single homogeneous highly permeable aquifer at a transboundary-scale. The typologies indicate different potential opportunities for management and sustainable development while also highlighting regions where mitigation is important to address depletion and degradation. Given that water quality degradation is likely to pose a greater threat than depletion for many of the typologies, systematic regular groundwater quality monitoring is fundamental to track changes and adopt appropriate management strategies. Strong scientific and management capacity at local levels, along with the legal and policy frameworks necessary to tailor management to local groundwater and surface-water conditions within typologies, will be essential in order to mitigate risks and take advantage of opportunities.


Polymethylmethacrylate (PMMA) also referred as (acrylic) bone cement is a non-degradable biomaterial that has been used in clinical orthopedic practice for several decades. PMMA can be used in a plain formulation, but is often used in an antibiotic-loaded formulation in (primary and revision) arthroplasty and in treatment of orthopedic infections as prosthetic joint infections (PJI) and chronic osteomyelitis. In treatment of PJIs antibiotic-loaded PMMA is often used as a carrier material for local antibiotic delivery in addition to treatment with systemic antibiotics. In this case, the antibiotic-loaded PMMA is often used as a spacer or as a bead chain. Since the introduction of PMMA as an antibiotic carrier there is a tremendous amount of scientific and clinical papers published, which studied numerous different aspects of antibiotic-loaded PMMA. This paper will review the research regarding basic principles of antibiotic-loaded PMMA as mechanism of action, antibiotic-release capacities, choice of antibiotics and influences on mechanical properties of PMMA. Subsequently, concerns regarding the application of antibiotic-loaded PMMA, biofilm formation, antibiotic resistance and local or systemic toxicity will be discussed. In addition to these subjects, the role of antibiotic loaded PMMA in clinical treatment of PJIs and chronic osteomyelitis is discussed in the final part of this paper. Bacterial infections are one of the most devastating complications in the field of orthopedic surgery. Especially infections related to implants e.g., cases of infection in total joint arthroplasty (TJA), or infections related orthopedic traumatology where implants are used for bone fixation are burdensome and difficult to treat. The infection rates vary based on the etiology of the infections, this review highlights the orthopedic prosthetic joint infections (PJIs) and trauma related chronic osteomyelitis. In cases of PJI the infection rates vary from <1% in primary total hip arthroplasty up to 10 to 14% in spine surgery or revision arthroplasty . In the The number of orthopedic implant surgeries is rising in the past years and is expected to increase in the upcoming years. Not solely the absolute numbers of PJIs are increasing, but also the relative infection rates are increasing . These nAs mentioned before, local antibiotic therapy is important in treatment of PJIs and chronic osteomyelitis. For the antibacterial effectiveness, it is important that the local antibiotic concentration exceed the minimal inhibitory concentrations (MIC) of the causative pathogens . DependiBone cement is used for almost 150 years since Gluck introduced it in 1870 for fixation of a total knee prosthesis . Back thIn the past decades, the optimization of antibiotic-loaded PMMA became an important research topic in the improvement of prophylaxis and treatment of PJIs . PMMA isin vitro studies showed that the pharmacokinetic release profiles of PMMA can be optimized by adjusting several properties of the PMMA. Since antibiotics are released by dissolution after contact with body fluids, an increase of the surface roughness and the porosity of the PMMA result in an increase of surface area, leading to an increased antibiotic release  release up to 93% of the added antibiotics  and achieve antibiotic concentrations up to seven times higher compared to normal PMMA beads  and the PROSTALAC® spacer . Some studies suggests that mobile spacers might even result in better infection treatment, although this is still under debate due to the absence of unequivocal evidence  and  joint mobility, which all favor the re-implantation of the definitive prosthesis during the second stage of PJI treatment, see l months . In contl months . In compl months . This isl months . Second-evidence . Disadvaevidence  and assoReduction of biofilms or prevention/inhibition of biofilm formation remains one of the major challenges in prevention and treatment of PJIs and chronic osteomyelitis. The phenomenon of biofilm formation is often termed “the race for the surface,” which was first described by Since bacteria in biofilm are protected by the biofilm layer they cause clinically relevant therapeutic and diagnostics challenges. The biofilm may cause difficulties in the microbiological diagnosis because the biofilm matrix can impede culturing of the bacteria, which can lead to false-negative cultures and missed diagnosis of infections. In order to prevent this there are many new developments in detaching the bacteria from their biofilm for better diagnosis .Therapeutic difficulties caused by biofilms are related to the decreased susceptibility for  antibiotics. The extracellular matrix of the biofilm causes an additional barrier resulting in decreased diffusion. In addition, substances of the matrix react with the antibiotics and thereby decrease the transport rates . Besidesin vitro studies showed that high local antibiotic concentrations inhibit, but not fully avoid, biofilm formation  antibiotics are the toxic side effects, especially cytotoxicity and systemic toxicity. Cytotoxicity after local antibiotic therapy is studied on cell survival and the capacity to recover of eukaryotic cells (like osteoblasts) in different ntamicin . For mosibiotics . AlthougAnother major concern in the application of antibiotic-loaded PMMA is the increase of bacterial antibiotic resistance. It is thought that after release of the majority of the antibiotics out of the antibiotic-loaded PMMA, the remaining antibiotics lead to sub-inhibitory concentrations, causing mutational antibiotic resistance . In addiWith the introduction of antibiotic-loaded PMMA around the 1970s and the specific gentamicin-loaded PMMA a few years later, the prevalence of PJIs decreased from around 5% to less than 1% . After sAntibiotic loaded PMMA spacers are used over two decades in two-stage PJI treatment with good clinical results. Different studies show that in treatment of infected THA as well in treatment of TKA, eradication rates of two-stage treatment are successful in 85 to 100% . HoweverAs PJIs and chronic osteomyelitis remain a substantial problem in orthopedic surgery, improvement of treatment of PJIs and chronic osteomyelitis with local antibiotic-loaded PMMA continues. Current research concepts are regarding surface modification and microencapsulation of specific antibiotics as e.g., rifampicin to enable biofilm breakdown and infection treatment by rifampicin loaded PMMA cement with good mechanical properties .In addition to development of antibiotic-loaded PMMA in prevention and treatment of PJIs, new carriers and coatings for controlled release of local antibacterial agents are important research topics for biofilm infections. Thereby, new carriers are interesting prophylactic opportunities for the uncemented prosthesis in TJA as well. These new strategies are based on adaptation of the surface of implants to prevent from infection, basic principles are (1) passive surface modification, (2) active surface modification and (3) local carriers/coatings, see In treatment of chronic osteomyelitis there is a tremendous increase in different biomaterials that might be suitable for local antibiotic delivery. In addition to local antibiotic delivery, these new biomaterials must have good bone defect filing capacities and should be biodegradable. Using these biodegradable local-antibiotic carriers enables the possibility for one-stage treatment of chronic osteomyelitis, which results in lower patient burden, less hospitalization and lower costs. In order to enable on-stage surgery these biomaterials must have proper biocompatibility to stimulate new bone formation and the must not have any toxic side effects on the surrounding tissues. Thereby they should have good biomechanical properties in order to prevent fractures or other mechanical failures after surgery. In the development of these biodegradable antibiotic-loaded biomaterials, different materials have been tested in pre-clinical and clinical studies. Examples of these materials are natural polymers like collagen and chitosan; synthetic polymers like polylactide; ceramics like calcium sulfates and calcium phosphates  and bioactive glasses . A recenTvV did the literature research, wrote the first draft, and finalized it to submitted version. JA and JG assisted with literature research, provided additional information, and reviewed the different versions.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


In the case of white wines, the comparison with REDOX-sensory scores showed that results from this modified DPPH assay are strongly correlated with sensory attributes . These results provide an unprecedented illustration of the important contribution of these sulfur-containing compounds to the radical quenching ability of white wines.The DPPH  assay is an easy and efficient method commonly used to determine the antioxidant capacity of many food matrices and beverages. In contrast with red wines, white wines are poorer in antioxidant polyphenolics, and the more hydrophilic sulfur-containing compounds in them may contribute significantly to their antioxidant capacity. The modification of the classical DPPH method, with a methanol-buffer and the measure of EC The aging of premium wines, and in particular dry white wines, has become an important scientific issue over the last decades, where the phenomenon called “premature aging” has appeared worldwide. In the context of sustainability, new processes based on the reduction of inputs, both at the vineyard and in the cellar (for instance sulfites), are sometimes considered as threats towards expected organoleptic optimums. Therefore, a better understanding of the physico-chemical mechanisms of oxidation is required to predict and control the appearance of premature aging. This is based in particular on the development of innovative predictive tools for determining the oxidative stability of a wine.Wine oxidative stability can be related to intrinsic and extrinsic factors that prevent aroma deterioration . Up to n50), which represents the quantity of the sample needed to decrease the initial concentration of the DPPH free radical by 50%. However, some studies showed the lack of correlation between the DPPH discoloration and the concentration of the substrate th), as follows:For the determination of the antioxidant capacity of wines, the molar number (or the concentration) of antioxidant compounds was unknown, therefore the molar ratio RnormV) to determine the EC20 of the wines.The absorbance was then normalized by the absorbance of a blank (water) and plotted against the corrected volume  were presented at room temperature in standardized black glasses coded with 3-digits following a Latin-square arrangement. For each sample, the panelists were first asked to rate the intensity of oxidation–reduction (REDOX-sensory score) orthonasally  using the protocol proposed by Ballester et al. 2018) [18 [20]. All measurements were performed in duplicate, and the results were expressed as mean ± standard deviation. Comparisons between methods (methanol and methanol-buffer) were carried out using the Student test, and the classification of antioxidant capacity of standard compounds was studied by ANOVA, followed by Tukey (HSD) test. The sensory results were analyzed using an ANOVA and PCA . These statistical analyses were done with the software XLSTAT. Figures were plotted using Matlab R2015a (MathWorks) and OriginPro 8 softwares .The modification of the classical DPPH method, with a methanol-buffer  provided new reference values for the antioxidant capacity of active compounds representative for white wines. In particular, and as a result of a significantly higher solubility in this methanol-buffer, sulfur-containing compounds such as cysteine, glutathione or methanethiol appeared to bear antioxidant capacity comparable to well known phenolic compounds, such as catechin, caffeic acid and ferulic acid, respectively. These results provide an unprecedented illustration of the important contribution of these sulfur-containing compounds to the radical quenching ability of white wines.The analysis of white wines from different vintages, and at different levels of oxidation, has further permitted validation of the method. Moreover, the comparison with REDOX-sensory scores, showed that results from this modified DPPH assay are strongly correlated with sensory attributes of the same wines. Therefore, such a modified DPPH method not only complies with requirements for the measurement of genuine antioxidant capacity of white wines, but also offers an interesting alternative for developing predictive tools of the ageing ability of white wines.


The study proposes a novel consensus strategy based on linear combinations of different docking scores to be used in the evaluation of virtual screening campaigns. The consensus models are generated by applying the recently proposed Enrichment Factor Optimization (EFO) method, which develops the linear equations by exhaustively combining the available docking scores and by optimizing the resulting enrichment factors. The performances of such a consensus strategy were evaluated by simulating the entire Directory of Useful Decoys (DUD datasets). In detail, the poses were initially generated by the PLANTS docking program and then rescored by ReScore+ with and without the minimization of the complexes. The so calculated scores were then used to generate the mentioned consensus models including two or three different scoring functions. The reliability of the generated models was assessed by a per target validation as performed by default by the EFO approach. The encouraging performances of the here proposed consensus strategy are emphasized by the average increase of the 17% in the Top 1% enrichment factor (EF) values when comparing the single best score with the linear combination of three scores. Specifically, kinases offer a truly convincing demonstration of the efficacy of the here proposed consensus strategy since their Top 1% EF average ranges from 6.4 when using the single best performing primary score to 23.5 when linearly combining scoring functions. The beneficial effects of this consensus approach are clearly noticeable even when considering the entire DUD datasets as evidenced by the area under the curve (AUC) averages revealing a 14% increase when combining three scores. The reached AUC values compare very well with those reported in literature by an extended set of recent benchmarking studies and the three-variable models afford the highest AUC average. Virtual screening (VS) involves different computational approaches aimed to identify from among huge molecular databases optimized sets of compounds which have the potential to bind a given biological target and which will undergo high throughput screenings (HTS) in order to identify novel hit compounds . VirtualOne of the simplest ways to evaluate the performances of a VS study is to measure how much the resulting datasets are enriched in active compounds by analyzing the so-called enrichment factors (EF) . SimilarEven though VS can also involve ligand-based approaches , dockEven though docking results are clearly influenced by the applied search algorithms, scoring functions represent the most critical factor in determining the overall reliability of docking simulations . On thesAs cited above, the procedures, in which different docking scores are combined, usually involve consensus strategies, and the choice of the optimal consensus algorithms plays a critical role in determining the overall VS performances . While lThe EFO approach can also be utilized when developing single variable models to quickly reveal the best performing scores while avoiding systematic analyses. In this case, the EFO algorithm does not involve optimization processes and thus the models and the corresponding EF values can comprise the entire DUD datasets without validation procedures. Here and in the following sections, the analyses will be subdivided into three parts which involve: (a) the primary Piecewise Linear Potential score (PLP) and its normalized values as directly computed by PLANTS; (b) the various scoring functions as computed by ReScore+ without post-docking minimization and (c) after post-docking minimization. While being less pronounced, the complex minimization also affords a beneficial contribution to the obtained performances as documented by the overall increase of 65% in the EF 1% averages when comparing the performances reached by rescoring minimized complexes with those obtained when using only the primary scores. When analyzing the specific results for each target class, the three parts reveal similar trends in which kinases are by far the most challenging targets. Serine proteases are the targets the performances of which reveal the greatest beneficial effect from the rescoring calculations while the EF values for nuclear hormone receptors appear to be poorly sensitive to the rescoring effects.On average, the similar trends observed for each target class regardless of rescoring and/or of complex minimization indicate that the obtained performances are similarly affected by the reliability of the generated poses, suggesting that rescoring analyses, while showing clearly beneficial effects, cannot completely elude the weaknesses of the utilized docking program.As seen for the EF average values, the EF enhancements are higher when considering the top of the rankings and decrease when extending the monitored rankings. Moreover, the EF enhancements significantly vary in the top of the rankings in the three study sections, while they are marginal and almost constant regardless of rescoring and/or minimization when considering larger parts of the rankings. The limited efficacy of the proposed consensus strategy when analyzing larger parts of the ranking can be explained by remembering that the EFO algorithm develops the linear equations by optimizing a cost function which, while also accounting for the distribution of the active compounds in the entire ranking, and is primarily focused on the number of active compounds within the first cluster. Since, the DUD datasets comprise on average ~ 3000 compounds see  and the When comparing the EF averages and the relative increases for the three parts of the study, one may notice the same trends already observed for the one-variable models with the scoring functions obtained by rescoring the minimized complexes which perform markedly better than the primary PLANTS scores. This observation suggests that this consensus strategy cannot upset the reliability of each single scoring function and thus the best models are conceivably generated by linearly combining the best performing scores. The limited improvements observed when using only the PLANTS scores can be explained by considering that the generated equations combine the various normalizations of the sole primary PLP score and thus the included variables, even when surpass the applied cross-correlation filter, comprise rather redundant information. In contrast, the other two parts of the study are based on conceptually diverse docking scores and thus clearly benefit from their linear combinations as seen in The analysis of the results obtained for the different target classes reveals trends very similar to those already seen for the one-variable models. Kinases remain the most challenging targets even when using consensus equations, while metalloenzymes and serine proteases show the highest EF averages. More importantly, the EF differences between one- and two-variable equations as computed for each target class see  reveal tBy considering the encouraging results provided by the VEGA-based scores, an additional study involved the development of two-variable equations by including only the VEGA-based scores computed after complex minimization. The study has the primary objective to evaluate in depth the recently proposed scores based on both the Molecular Lipophilicity Potentials   those produced by machine learning (ML) techniques.When considering the various classes of targets, the here proposed consensus approach yields the best performances in all considered classes apart from kinases where the ML study reports markedly better results. This finding is in agreement with the already discussed difficulty in screening the kinase datasets and suggests the opportunity of testing diverse approaches in order to find the best performing strategy. Thus, the here reported method affords the best model in 19 out of 40 datasets, the same result is reached by the ML approach, while in 2 cases the best results are provided by the mRAISE method. Finally, http://dud.docking.org/). All utilized molecules (protein targets and ligand datasets) were downloaded from the DUD website, while the ligand structures were utilized as they were, the protein structures underwent a preliminary set-up before docking simulations. In detail, the resolved complexes were assembled and completed by adding the non-polar hydrogen atoms using VEGA  [As mentioned in the Introduction, the present study involved all the 40 datasets included in the original version of DUD (see d files) . The so MAX-MIN ant colony optimization (ACO) [All docking simulations were performed by using PLANTS  which geon (ACO) . In detaon (ACO) . All menon (ACO) , applyinon (ACO) . In detaon (ACO) , the Lenon (ACO) , the numon (ACO) , the thron (ACO) , and theon (ACO) . As discussed in the Introduction, the primary objective of the study involves the assessment of suitably optimized linear combinations of different scoring functions as a consensus strategy for evaluating virtual screening campaigns. The selected linear equations were computed by using the EFO classification algorithm, an approach recently proposed to analyze unbalanced datasets and which can find in virtual screening campaigns a fruitful applicative field . BrieflyAll generated models with two and three variables were evaluated and screened by a per target validation using the procedure implemented by default in the EFO algorithm. Indeed and as previously detailed, the EFO algorithm automatically subdivides the input dataset in training (80%) and test (20%) sets, develops the models using the training set and validates by applying them to the training set. Moreover and to minimize the randomness, this validation task is repeated five times and the models are evaluated and prioritized by considering their average performances as obtained during the validation phase. The study describes the exploitation of linear combinations of more than one docking score as a consensus strategy to enhance the reliability of docking simulations in virtual screening campaigns. Indeed and even though many consensus approaches have been proposed in the last few years, including protocols based on machine learning techniques, the (seemingly simple) linear combinations of scoring functions have received little attention in this field. Here, the consensus linear equations are developed by using the EFO classification method which was recently proposed to analyze highly unbalanced datasets and which can find really fruitful applications for evaluating virtual screening campaigns.Clearly, an extensive assessment of the performances offered by such a consensus strategy would require further analyses considering (a) different and more challenging datasets, (b) diverse docking programs, (c) additional scoring functions also including molecular descriptors, and (d) more than one computed pose per ligand to be treated in the framework of the recently proposed binding space. That being said, the above-described results provide a convincing confirmation of the remarkable performances of the proposed consensus strategy and allow some general rules to be derived: (1) the linear combination of two or three docking scores represents a satisfactory balance between performances and calculation costs; (2) while clearly depending on the reliability of the computed poses, rescoring calculations appear to be a powerful and straightforward strategy to enhance the performances of a docking simulation; (3) post-docking complex minimization exerts marked beneficial roles especially when considering non-primary scoring functions. As an aside, the study comprises an extensive and successful validation of the reliability of the MLP iteraction scores and the score based on the number of contacts in the field of virtual screening simulations.


This study explores the effect of slum rehabilitation on appliance ownership and its implications on residential electricity demand. The low-income scenario makes it unique because the entire proposition is based on the importance of non-income drivers of appliance ownership that includes effects of changing the built environment (BE), household practices (HP) and appliances characteristics (AC). This study demonstrates quantitatively that non-income factors around energy practices influence appliance ownership, and therefore electricity consumption. The methodology consists of questionnaire design across the dimension of BE, HP and AC based on social practice theory, surveying of 1224 households and empirical analysis using covariance-based structural equation modelling. Results show that higher appliance ownership in the slum rehabilitation housing is due to change in household practice, built environment and affordability criteria of the appliances. Change in HP shifts necessary activities like cooking, washing and cleaning from outdoor to indoor spaces that positively and significantly influences higher appliance ownership. Poor BE conditions about indoor air quality, thermal comfort and hygiene; and product cost, discounts and ease of use of the appliances also triggers higher appliance ownership. The findings of this study can aid in designing better regulatory and energy efficiency policies for low-income settlements. •Slum rehabilitation causes household practices to shift indoors.•Shifting of household practices to indoors increase appliance ownership.•Slum rehabilitation increases energy intensive activities.•Non-income factors around energy practices influence appliance ownership. The International Energy Agency (IEA) states that OECD countries account for 65 per cent of the total residential electricity consumption globally, while electricity demand in non-OECD countries, especially in the Global South, have grown at twice the rate compared to the OECD countries . It is eEveryday activities of occupants shape their need for household appliances which influence energy consumption. These activities represent practices, which are routinised type of behaviour of normal life . These pKnown drivers of residential energy use at a macro-level are income, climate, demographic characteristics, along with energy price dynamics, dwelling type and technology evolution including information and communication technologies (ICT) , Rao an, Ekholm. In IndiIn this study, we investigate whether the change of the built environment of slum dwellers to slum rehabilitation housing (SRH) affect their residential electricity use. In doing so, we investigate the number of appliance purchase in the slum rehabilitation housing after shifting and assume it as an indicator of end-user demand for residential electricity. This system boundary defines the novelty of this work, where we link change in built environment, household practice and appliance characteristics as essential drivers of appliance ownership in low-income settlements.The policy of ‘Slum Rehabilitation Housing’ was adopted in 1995 by the Government of Maharashtra as a response to redevelop the slums into high-rise social housing by incentivising the private sector to participate in the redevelopment of slum communities. The process mandated to re-house the slum dwellers onsite in high-rise buildings that includes legal entitlement to a stipulated 269-square-foot apartment, including a bathroom with tap water and a kitchenette. This policy provided the slum dweller access to a cross-subsidised, free of cost house, without burdening their time or economic poverty . The houWe base our study in an SRH society in Mumbai that comprises of occupants who have been living here for more than six months. Previous studies have shown that these SRHs often resemble ‘vertical slums’ characterised through the hyper-density of occupants, sub-standard dwelling quality and inadequacy to provide basic amenities . These sThe primary hypothesis of this study is that higher appliance ownership in the slum rehabilitation housing is due to change in household practice triggered by the change in the built environment from the horizontal slums to SRHs. Six sub-hypotheses  to H6 we2Appliance ownership has mainly been discussed or predicted using empirical research based on societal trends, while very few studies examined determinants of appliance ownership at a household level . MacroecAmong the few studies of household level determinants, most of the studies have reported household size to be a significant non-income determinant, along with housing type and age . In ruraA significant effect of family composition  and the age of the household responsible person (HRP) has been widely acknowledged in the literature as a crucial non-income driver of energy consumption . HoweverOther commonly discussed built-environment elements that influence energy consumption are the number of rooms, bedrooms, floors and the floor area in addition to the presence of electric space heating, ventilation, electric hot water heating systems and air-conditioning systems . As mentIn another study by While these drivers represent the material aspect of Social Practice Theory (SPT), the meaning and skills represent the abstract element of the occupants' socio-cultural dynamics that drives their energy stacking behaviour. In a study by Rao and Ummel's , Bisaga 33.1Mumbai has an estimated population of 12.4 million as per 2011 Census data, out of whom 42 per cent (i.e. 5.2 million) of the population lives in the slums . Since tThe surveys spanned across 1,224 households which were selected using a stratified random sampling method across the four SRHs. A survey questionnaire was developed comprising of three segments that recorded the change in household practices on moving to SRH from the horizontal slums, perception of built environment and household appliance ownership criteria. The questions on the change in household practices were drawn after the work of  was included to understand the gender dynamics in the appliance-purchase decision. The interviewees were asked, ‘Who usually makes the appliance purchase decisions?‘. Other variables in this segment included documentation of the time-related mandatory household activities. Further narratives were collected based on the methodology of . Similar scale was used to report the occupant's perception of the thermal comfort in their present rehabilitation houses . Additionally, an ordinal variable was added to report the change in thermal comfort conditions as compared to the horizontal slums from the occupant's perspective and was coded as ‘Same= 0; Less comfortable = 1; More comfortable = 2’.Questions relating to the built-environmental attributes were derived from the recent study of  defining product characteristics was used that included size, brand, product cost, quality, availability of discount and the ease of using the appliance  was conducted to identify the strength of each path of the hypothetical model see  and veriSix hypotheses were formulated based on the evidence from the literature on social practice and residential electricity use, built environment and its effect on energy demand and appliance ownership characteristics in low-income tenement housing . It remains a salient feature of this study that the effect of low-income built environment transition can influence household electricity demand through a change of household practices. The typical appliance layout of horizontal slums and SRH apartment units are illustrated in The current status of appliance ownership is illustrated in Based on While almost 99% of the surveyed households had ceiling fans see , there wThe cleaning time also reduced as the occupants had to cater to their own 265 square feet rooms rather than cleaning several slum houses and the collective social spaces as in the horizontal slums. This shifting of household practices to private and indoor life in the SRH due to the architectural design of the built environment encouraged adoption of energy intensive appliances like washing machines and refrigerators. It is discussed in detail in section A stark difference is in the time spent in subsistence activities see d, wherei4.2The final SEM model and model fit indices for all data (n = 1224) are shown in Non-income factors chosen for this study was represented through six hypotheses illustrated in Specific appliance characteristics that have a significant positive effect on the household practices of the occupants are brand , product cost , quality , discount available  and ease of use  see . As mentThe change of the built environment influences changes in household practices see , and it Model results show that there is a significant increase in indoor household activities upon moving into the SRH that significantly influences the household practices  that in turn strongly influences higher appliance ownership  see . IncreasThe hypothesis Considering the direct and indirect effects on the appliance purchase after shifting, the total estimated effect of each factor is shown in 4.3The empirical results presented in section bazaar’ dismissed the need for refrigerators. The ineffective design of SRH with low accessibility made the activities singular and space-bound, triggering energy-intensive indoor living. Presently, refrigerator and TV are the most common appliance that was purchased after shifting to the SRH  through the Gates-Cambridge Scholarship. This study is in parts funded by Ministry of Human Resource Development, Government of India (Grant no. 14MHRD005) under the Frontier Areas in Science and Technology grant awarded to RB.RD is supported by


Spontaneous flexor tendon rupture is rare, occurring most commonly in the little finger or flexor pollicis longus. To the best of our knowledge, there have been no reports of spontaneous flexor tendon rupture due to primary distal radioulnar joint (DRUJ) osteoarthritis (OA). We present a case of spontaneous flexor tendon rupture in the index finger due to primary DRUJ OA in a 71-year-old female farmer. Surgical exploration confirmed that, at the wrist joint level, the flexor digitorum profundus of the index finger had undergone degeneration and complete rupture. The flexor digitorum superficialis of the index finger was elongated and thinned. A bony spur toward the volar side was covered with synovial fluid from a pinhole-sized perforation of the capsule. The combination of direct friction from the DRUJ spur and the matrix metalloproteinases in the synovial fluid from the perforation of the DRUJ capsule may have caused the spontaneous flexor tendon rupture. Palmar-side symptoms associated with DRUJ OA should be carefully examined because of the risk of spontaneous flexor tendon rupture. Spontaneous flexor tendon rupture is relatively uncommon and is usually caused by trauma, inflammatory disease, steroids , or surgical complications from plate, and carpal bone and joint disorders , 2. TherThe study protocol adhered to the ethical guidelines of the 1975 Declaration of Helsinki, and the study was approved by the institutional review broad of our institute. The patient was informed that this case study would be submitted for publication, and she provided informed consent.A 71-year-old woman was prescribed analgesics at another orthopedic clinic because of pain and swelling in the right carpal area. Half a month later, she could not flex the index finger of her right hand. She had no past history of trauma, carpal bone and joint disorders, or inflammatory disease and had not taken any steroid injection recently. She has been a farmer for a long time. On clinical examination, she was not able to actively flex the distal interphalangeal joint of her index finger. The proximal interphalangeal joint could be flexed to 40°. The anterior-posterior and lateral plain radiographs showed a bony spur arising from the volar ulnar aspect of the distal radius Figures . ComputeDuring surgery, under general anesthesia and using tourniquet control, a zig-zag incision was made at the level of the DRUJ on the palmar side. Surgical exploration confirmed that at the wrist joint level, the flexor digitorum profundus (FDP) of the index finger had undergone degeneration and complete rupture. The flexor digitorum superficialis (FDS) of the index finger was elongated and thinned. The FDP of the middle finger had undergone slight degeneration; however, tension of the FDP of the middle finger was normal . The bonDRUJ OA occurs as the result of a variety of mechanisms, including primary lesions, inflammatory arthritis (particularly rheumatoid arthritis), posttraumatic causes , and congenital or developmental abnormalities of the joint surfaces  , 11. TheThe most common complication of DRUJ OA is extensor tendon rupture . NeverthMost reports of spontaneous extensor tendon rupture due to DRUJ OA may have involved perforation of the DRUJ capsule , 12, 18.In conclusion, DRUJ OA is a rare cause of flexor tendon rupture and attrition. Attention should be paid when evaluating a patient with palmar-side symptoms of the wrist joint with DRUJ OA because of a risk of spontaneous flexor tendon rupture.


To the best of our knowledge, only 2 cases of Creutzfeldt-Jakob disease demonstrating nigrostriatal dopaminergic deficits in vivo using DaTSCAN have been published in the literature.Sporadic Creutzfeldt-Jakob disease (sCJD) is a type of progressive, subacute encephalopathy associated with spongiform degeneration of the central nervous system. sCJD includes a broad and heterogeneous spectrum of clinical variants, but extrapyramidal symptoms and signs at disease onset were rarely reported. We describe a case of unilateral parkinsonism associated with pathological  Parkinsonism as the first symptom of sporadic Creutzfeldt-Jakob disease (sCJD) is rarely described –3. The aWe present the case of a patient with sCJD manifesting with gait abnormality and right-side bradykinesia and hypokinesia, associated with presynaptic dopaminergic deficits. Our case supports the hypothesis of nigrostriatal pathway alterations in CJD.123I-ioflupane SPECT (DaTSCAN), which revealed asymmetrical tracer uptake in the caudate nuclei (decreased uptake in the left caudate nucleus) and a near-complete lack of activity in the putamina, particularly on the left side . The examination revealed no further abnormalities. With the clinical suspicion of parkinsonism, we performed a complete blood test performed at baseline, with either normal or negative. A brain MRI scan revealed increased signal intensity in the left putamen with diffusion restriction, suggesting lacunar infarction . By the eft side . On disceft side . PRNP gene; the study of codon 129 polymorphisms revealed methionine/valine heterozygosity (M129V). The probable diagnosis of sporadic CJD was reached based on the World Health Organization (WHO) diagnostic criteria iodophenyl) nortropane. This cocaine analogue has a high binding affinity for and acceptable sensitivity to dopamine transporters (DAT), making it an excellent marker of viability and function of nigrostriatal neurons. Numerous studies of Parkinson's disease have evaluated the use of this technique to confirm clinical diagnosis. In the context of Creutzfeldt-Jakob disease, in contrast, DaTSCAN may be useful for in vivo confirmation of the presence of striatal dopaminergic degeneration.Further, lesions to the nigrostriatal pathway have never been studied in detail . To our Although our patient was admitted due to pseudoictal symptoms of language impairment, his gait disorder had appeared a month previously. In our case, the DaTSCAN revealed abnormal tracer uptake in the left caudate nucleus and reduced tracer uptake in the putamina ; these findings were coherent with the patient's symptoms of right-sided bradykinesia. It may be argued, however, that the patient had mixed parkinsonism  based on functional neuroimaging results and the presence of a lesion to the left putamen on diffusion-weighted MRI sequences; this lesion may reflect the spongiform changes typical of sCJD. These findings were consistent with anatomical pathology results from the postmortem studies, which revealed dopaminergic neuron loss in the substantia nigra, caudate nucleus, and putamen . This suIn conclusion, parkinsonism in CJD may be caused by lesions to the basal ganglia affecting the lenticular nucleus, or by nigrostriatal pathway degeneration. Much remains to be understood about DaTSCAN as an in vivo functional technique for the study of basal ganglia motor circuits and their role in the pathogenesis of this symptom in CJD. In any case, DaTSCAN seems to be a promising, noninvasive method for identifying lesions to the dopaminergic pathway; combined with MRI, it may help in the diagnosis of CJD.


Delivery of compounds to the peripheral nervous system has the potential to be used as a treatment for a broad range of conditions and applications, including neuropathic pain, regional anesthesia, traumatic nerve injury, and inherited and inflammatory neuropathies. However, efficient delivery of therapeutic doses can be difficult to achieve due to peripheral neuroanatomy and the restrictiveness of the blood-nerve barrier. Depending on the underlying integrity of the blood-nerve barrier in the application at hand, several strategies can be employed to navigate the peripheral nerve architecture and facilitate targeted delivery to the peripheral nerve. This review describes different applications where targeted delivery to the peripheral nervous system is desired, the challenges that the blood-nerve barrier poses in each application, and bioengineering strategies that can facilitate delivery in each application. Delivery of therapeutic compounds to the peripheral nervous system (PNS) is difficult to achieve due to complexities of the peripheral neuroanatomy and the restrictiveness of the blood-nerve barrier (BNB). The endothelial cells that line the endoneurial vasculature are non-fenestrated and linked by specialized tight junctions, forming a restrictive barrier that protects the endoneurial microenvironment  and ensuWhile the BNB is fundamental for normal function it hinders the transport of therapeutic compounds from the vasculature into the nerve. The sciatic nerve itself is impermeable to hydrophilic small molecules  as well There are several scenarios where targeted delivery to the peripheral nerves is desired, and the features of BNB dictate the choice of an appropriate drug delivery strategy Figure . For exaLocal, controlled delivery at the site of repair also allows for administration of an experimental therapy that may otherwise be contraindicated as a systemic treatment . MechaniIn another example, local, controlled delivery of anesthetics or analgesics to peripheral nerves is necessary to provide a nerve block or relieve neuropathic pain. Here, the perineural and endoneurial barriers are intact and restrictive. Local anesthesia bears several advantages over general anesthesia for surgical procedures of the limbs , and local peripheral nerve blocks can also be used to treat chronic neuropathic pain . StrategStrategies based in gene therapy can also be exploited to target peripheral nerves in the presence of an intact barrier. Here, targeting can be conveyed by incorporating bacterial toxins that display natural neurotropism for ganglioside receptors that are enriched at presynaptic terminals. Retrograde transport can facilitate delivery of encapsulated cargo along the axon, to the dorsal root ganglia, or spinal cord. This approach has been investigated in models of regeneration , inheritIn each of the aforementioned scenarios, the BNB poses a challenge in the delivery of therapeutic cargo  to the endoneurial compartment. In this review, we describe bioengineering strategies that can be used for therapeutic, targeted delivery to the peripheral nerve. These strategies  allows for direct application of the nerve conduit or graft. Both clinically and experimentally, biomaterials incorporated into nerve conduits facilitate controlled release and protection of protein cargo, while providing structural support to regenerating axons.The restrictiveness of the BNB is not a primary matter of concern when considering delivery after injury or trauma, as these events are associated with subsequent disruption of the BNB . In addiBioengineered nerve conduits can be synthesized from a broad range of natural [e.g., collagen , lamininWhile the basic structure of a bioengineered nerve conduit is a hollow tube, several strategies can be employed to incorporate cargo resulting in tunable release kinetics . DiffereEach of the different methods for incorporating cargo into an engineered nerve conduit described above is associated with different release mechanisms and release kinetics. When a growth factor is simply in solution in the lumen of the conduit, release will occur via diffusion at a rate more rapid than desired for most applications. Incorporation of growth factor into degradable microspheres typically results in an initial burst release from the surface, followed by sustained release as the polymer breaks down and releases the encapsulated payload. Depending on the desired application, the release kinetics can be further tuned by modifying polymer composition, producing layered microspheres or conduits  or co-enin vivo  followed by NGF . In thisin vitro  is an immunosuppressant used in conjunction with cyclosporin A following organ transplantation that has several non-immune effects attributed to its use. Early studies demonstrated that FK506 promotes neurite outgrowth in vitro  and enha Figure . Nerve iA strategy to advance peripheral anesthesia and pain control methods incorporates the use of stimulus-responsive biomaterials. Such a delivery system would release minimal drug in the absence of an external stimulus, allowing for on-demand adjustment of the local dose of anesthesia according to the patient’s needs. Several methods of external triggering can be engineered into delivery systems to facilitate on-demand delivery of a payload, including near infrared (NIR) light  and ultrUltrasound has recently been investigated as a trigger for an on-demand system to release local anesthetic. An attractive feature of this system is that ultrasound is already incorporated into clinical practice to guide the placement of epi- and intraneural injections of local anesthetics . Rwei etThe same investigators have also explored local anesthetic delivery systems that are triggered by near-infrared light , 2017b. An alternative to the application of potentially toxic local anesthetics for the management of pain or production of sustained nerve block is the use of endogenous, hydrophilic opioid peptides. Mu-opioid receptors are expressed along the axons of peripheral nerves and are upregulated after injury or inflammation . While mSeveral strategies exist to disrupt the perineural and endoneurial barriers and facilitate targeted delivery of hydrophilic compounds. One strategy extends earlier findings that transient opening of the blood-brain barrier can be achieved through infusion of a hypertonic solution . InitialFollowing the determination that the barrier-opening effects of hypertonic saline were mediated through claudin-1, subsequent studies demonstrated that perineural application of a claudin-1 peptidomimetic facilitated barrier opening and transport of hydrophilic analgesics to axonal nociceptors in normal rats . Furtherin vivo or clinical application due to toxic, inflammatory, and immunogenic complications and safety concerns.One strategy that confers targeted delivery to the PNS despite the presence of an intact BNB employs basic principles of gene therapy. Initial approaches to nerve-targeted gene therapy exploited the natural neurotropism exhibited by many viral vectors, such as the affinity of herpes simplex virus for peripheral sensory neurons of the DRG . While rSeveral natural toxins exist that bind to receptors located within the central and peripheral nervous systems and could be used to engineer a targeted delivery system . TargeteTetanus toxins are clostridial toxins that exhibit high affinity binding to ganglioside receptors, primarily the ganglioside GT1b receptor, on the presynaptic terminals of peripheral nerves . This hiin vitro (in vivo uptake and retrograde transport of Tet1-conjugated green fluorescent protein (GFP) to the motor neuron cell body from the site of injection (lumborum of latissimus dorsi) (N-(2-hydroxypropyl) methacrylamide (HPMA) and oligolysine/Tet1 macromonomers that formed complexes with plasmid DNA. In this system, Tet1 serves as the neuron-specific targeting entity, oligolysine serves to bind and protect nucleic acid, and HPMA provides stability. The authors demonstrated successful gene delivery to neurons in vitro (E-caprolactone)-block-poly(ethylene glycol) (PEG-b-PCL) have also been demonstrated to deliver nucleic acids to central nervous system (CNS) neurons when utilizing Tet1 as a targeting moiety  . To testin vitro . Complexg moiety .While Tet1 has been incorporated as a targeting moiety into neuron-specific delivery systems, further advances have been made with inactive, nontoxic mutants or fragments of the tetanus toxin holoenzyme. in vivo nerve crush injury model  which displays a natural neurotropism through its binding to the ganglioside GM1. The specificity of CTB for GM1 has recently been exploited to target mesoporous silicone nanoparticles (MSNPs) to motor neurons . This sy in vivo , and demin vivo. Future successful delivery systems may combine aspects of each strategy to facilitate coordinated release of multiple factors. The emergence of these studies in just the past year or two suggests that this field will be rapidly expanding.The next wave of neuron-targeted delivery strategies that exploit mechanisms of retrograde trafficking lies in delivery of diverse cargo that facilitates functional improvement in different disease models. In this regard, the payload capacity of mesoporous silicon nanoparticles may serve as a distinct advantage over cationic nanocomplexes, enabling the delivery of small molecules and proteins along the axon. However, as demonstrated by Lopes et al., delivery of plasmid DNA encoding a neurotrophic factor can result in increased protein expression and functional protection Despite advances in bioengineering to deliver compounds to the peripheral nerve as described above, targeted delivery for treatment of diseases of the PNS remain elusive. The sheer length of peripheral nerves may necessitate systemic delivery, while the selectivity of the BNB restricts access of systemically administered agents to the cells of the PNS. The most promising strategies will incorporate PNS-specific modifications to systemically administered nanocarriers, such that the administered compounds will (I) be protected from degradation/phagocytosis/opsonization within the circulation, (II) identify and recognize the targeted peripheral nerves, and (III) be able to cross the BNB.In vitro experiments demonstrated that fluorescently labeled DOPC liposomes were internalized by Schwann cells, sensory neurons, and motor axons, but not skeletal muscle cells. When administered intravenously, DOPC liposomes were detected at elevated levels in sciatic nerve Schwann cells as well as in highly vascularized regions of the brain. Detection of DOPC liposomes in the liver and kidney indicated clearance through these routes; however, collectively the data suggest that modified liposomes do cross the BNB and are taken up by myelinated peripheral nerves.A novel preparation of dioleoyl-phosphatidylcholine (DOPC) based liposomes preferentially targeted peripheral neurons and Schwann cells when conferred with some key modifications that mimic properties of neural cell membranes. Investigators added Poloxamer 188, a nonionic emulsifier that has been demonstrated to facilitate transport across the blood brain barrier, and supplemented liposomes with cholesterol, which is expressed at high levels in neuronal cells, especially myelinating glia . In vitrWhile liposomal preparations may confer specificity to neurons, it may be necessary for a delivery system to target PNS-specific receptors expressed by BNB endothelial cells, such that they may be accessed from the circulation. Several strategies have demonstrated delivery of cargo across the blood-brain barrier by targeting CNS-specific endothelial receptors, a situation analogous to that encountered in the periphery. The localized expression pattern of transporters and receptors to brain endothelial cells facilitates carrier-mediated transport of circulating particles with a surface modified to express the ligand . Deliverin vitro cultures -1 by endoneurial endothelial cells during inflammation has been observed in vitro  and in v(in vivo . Nanopar(in vivo . LFA-1-pin vitro . Similarin vitro . While iications , ICAM-1 Bioengineering has fueled advances in delivery of therapeutic cargo to the PNS, which can be difficult due to the presence of the BNB. The strategies that will be incorporated by a drug delivery system are dictated by the status of the BNB during injury, disease, or homeostatic conditions. These diverse strategies include coordinated release of multiple growth factors at the site of injury to facilitate repair in the presence of a breached barrier, or transient, localized barrier permeabilization and on-demand release to deliver peripheral nerve anesthesia or analgesia. In addition, the neurotropism of certain bacterial toxins can be exploited to deliver a customizable array of encapsulated cargo to the peripheral nerve via retrograde axonal transport for treatment of inherited neuropathies, neuropathic pain, and nerve injury. Finally, methods of targeting the BNB from the systemic circulation may include specific formulations of liposomes and surface functionalization with endothelial ligands. While much progress has been made, future clinical applications of targeted delivery to the peripheral nerve rely on continued advancement of biomaterials-based drug delivery systems, as well as further elucidation of the molecular characteristics of the BNB during health and disease.KL conceptualized the topic and prepared the first draft of the article. EB revised all sections critically. Both authors read and approved the final submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.