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Year : 2014 | Volume : 7 | Issue : 2 | Page : 93--103
Heat shock proteins and parasitic diseases: Part 1: Helminths
Sherif M Abaza
Department of Parasitology, Faculty of Medicine, Suez Canal University, Ismailia, Egypt
Department of Parasitology, Faculty of Medicine, Suez Canal University, Ismailia
Heat shock proteins (HSPs) are highly conserved and immunogenic proteins that are shared among diverse groups of mammals and microbial agents. They are categorized into different families according to their molecular weight. HSPs are involved in a variety of cellular processes and essential to cell survival. They are also implicated in immune pathology and clinical manifestations of a variety of autoimmune diseases and/or metabolic disorders such as atherosclerosis, diabetes and systemic lupus erythematosus. Their role in antigen cross-presentation and cancer immunotherapy as well as initiators of immune response and targets of autoimmune attack was also reported. The objectives of the current presentation are to summarize the functional properties of HSPs and their role in innate and acquired immune responses, to throw light on their role in pathogenesis and parasites survival, to review the literature searching for new drug discovery and vaccine candidates for parasitic diseases, and finally to present their use in diagnosis and genotyping of some parasitic diseases. Heat shock proteins (HSPs) are highly conserved and immunogenic proteins that are shared CONTENTS Introduction 1. Functional Properties of HSPs 1.1. Innate immunity 1.2. Adaptive immunity: 1.3. HSPs as cancer vaccines: 1.4. HSPs as infectious disease vaccines 1.5. HSPs and apoptosis 2. Heat Shock Proteins and Helminthes 2.1. Schistosoma spp. 2.2. Echinococcus spp. 2.3. Strongyloides spp. 2.4. Trichinella spiralis 2.5. Filarial nematodes 2.6. Other helminthes Concluding Remarks References Abbreviations: APC: Antigen-presenting cell; CTL: Cytotoxic T-lymphocyte; E/S: Excretory/secretory; gp96: a member of HSP90 family; GST: Glutathione-S-transferase; HC: Hydatid cyst; HSP: Heat shock protein; IFN: Interferon; IL: Interleukin; MHC: major histocompatibility complex; NK: Natural killer; SEA: Soluble egg antigen; TLR: Toll-like receptor; TGF: Transforming growth factor; TNF: Tumor necrosis factor.
Abaza SM. Heat shock proteins and parasitic diseases: Part 1: Helminths.Parasitol United J 2014;7:93-103
Abaza SM. Heat shock proteins and parasitic diseases: Part 1: Helminths. Parasitol United J [serial online] 2014 [cited 2023 Feb 1 ];7:93-103
Available from: http://www.new.puj.eg.net/text.asp?2014/7/2/93/149556
Stress proteins or heat-shock proteins (HSPs) are among the highly conserved and immunogenic proteins shared among diverse groups of mammals and microbial agents [1] . HSPs were first recognized when the temperature of an incubator housing Drosophila was elevated, resulting in a change in the pattern of chromosomal puffing within the salivary glands [2] . In addition to heat, these proteins were found to be inducible upon exposure to a range of environmental stresses including oxygen deprivation, pH extremes, and nutrient deprivation [1] . HSP synthesis occurs at 5-15°C above the optimal organism temperature depending on its growth temperature range [3] . Generally, there is a transient increase in HSPs synthesis at a low level of temperature elevation, with a more sustained response observed at higher temperatures, a pattern of response observed in numerous organisms. For example, heat shock of BCG by increasing the temperature to 42°C results in the production of both HSP65 and HSP70, whereas HSP70 synthesis is more pronounced at 45°C. The investigators added that both heat-shock response and upregulation of HSPs are observed in all tissues and in both prokaryotic and eukaryotic organisms, indicating that it is a ubiquitous and critical biological response [4] .
These proteins have been categorized into different families according to their molecular weight, for example HSP110, HSP90, HSP70, HSP60, HSP40, HSP20-30, and HSP10 [5],[6] . For uniformity, Kampinga et al. [7] proposed guidelines for the nomenclature of various human HSP families. However, HSPs could be categorized into different functional systems, with some overlap. The HSP60-HSP10 system is involved in classical protein folding, whereas the HSP70-HSP40 system stabilizes peptides in a linear, unfolded state and delivers them to the HSP60-HSP10 system [8] . The HSP90 family is found predominantly in the cytoplasm and is believed to mediate the folding of specialized proteins such as steroid receptors and protein kinases [9] . Thermal tolerance, disaggregation, and unfolding of aggregated proteins for enzymatic digestion are handled by the larger HSP100 [10] . Being involved in such a variety of cellular processes, it is not surprising that the majority of HSPs (HSP60, HSP70, and HSP90) are fundamental to cell survival, and mutation or deletion of the major HSP which is often lethal to both cells and organisms. In addition, HSP70 plays important cellular roles in protein synthesis and modifying processes, and can combine with many kinds of antigenic peptides to form antigen peptide-HSP compounds [11] . Furthermore, several studies have shown the ability of HSP70 to target vaccine antigens to antigen-presenting cells (APCs), including dendritic cells and macrophages, increasing vaccine immunogenicity [12],[13],[14] . Some HSPs were implicated in immune pathology and clinical manifestations of a variety of autoimmune diseases and/or metabolic disorders such as atherosclerosis [15] , diabetes [16] , systemic lupus erythematosus [17] , and Behcet's disease [18] . The role of some HSPs in antigen cross-presentation [5] and cancer immunotherapy [19] has also been reported. In addition, these proteins are initiators of immune response as well as targets of autoimmune attack. That is Microbial HSPs may stimulate cellular or humoral immune responses that might be cross-reactive with the corresponding self-HSPs or other self-antigens, leading to the induction of autoimmunity [6] .
HSPs are also known to play critical roles in innate immunity as well as adaptive immunity. They are capable of: (a) Activating specific toll-like receptors (TLRs), (b) Inducing the maturation as well as influencing the activation of APCs, (c) Providing polypeptides for specific triggering of the acquired immune response, and (d) Playing a major role in cross-presentation of extracellular antigens (e.g. microbial antigens) in parenchymal cells by the class I pathway, resulting in the induction of CD8+cytotoxic T-lymphocyte responses [20],[21].
Besides, HSPs may facilitate the regulation of effector responses under appropriate conditions, which could be achieved by increasing the production of anti-inflammatory cytokines so as to deviate the cytokine balance from a proinflammatory type to an anti-inflammatory type [22] . HSPs may induce different types of regulatory T cells, including CD4 + Tr1 cells that secrete interleukin-10 (IL-10), and CD4 + CD25 + , and the transcription factor Forkhead-box-P3 (Foxp3)-expressing Treg that produce transforming growth factor-β (TGF-β) and IL-10 [5],[16],[19] .
It was shown that ATPase activity is essentially associated with the functions of the major HSP families. In addition to ADP/ATP, the peptide binding of the HSP90 system is under the control of calcium levels that induces the required conformational changes for peptide binding [9] . However, it was argued that HSPs constitute a third functional group of adjuvants [23] as they were recognized as major immunogens against pathogens [5] . It was shown that apart from acting as immunogenic antigens themselves, HSPs can also act as adjuvants to stimulate the immunogenicity of heterologous polypeptides to which they are either covalently or noncovalently coupled [24] .
The interactions between HSPs and parasitic diseases were reviewed in three articles. Shonhai et al. [25] focused on intracellular protozoan parasites such as Leishmania spp., Tryapnosoma cruzi, Plasmodium falciparum, and Toxoplasma gondii, and reviewed the role played by HSP90 in the development and growth of all of these intracellular parasites, as well as the biological functions of HSP70-HSP40 interactions. A year later, two review articles were published by Indian scientists focusing on HSP90. The potential importance of HSP90 in the growth and pathogenesis of several parasites such as Giardia, Plasmodium, Trypanosoma, and Leishmania was pointed out [26],[27] . Thus, according to their view, several HSP90 inhibitors are expected to be promising targets as new drugs and candidate vaccines.
The objectives of the current review are to: (a) Summarize the functional properties of HSPs and their role in innate and acquired immune responses, (b) Shed light on their role in the pathogenesis and survival of the majority of parasites, (c) Review the literature for new drug discovery (HSPs-specific inhibitors) and vaccine candidates for parasitic diseases, and (d) Present other considerable uses of these biomarkers in parasitic diseases.
Functional properties of heat-shock proteins
The highly conserved HSPs molecules are defined as molecular chaperones. Their main function is the correction of the folding or functional conformation of cell proteins (under stress). This correction improves the capability of the cell proteins to: (a) Achieve their native form, (b) Be directed to the functional location (e.g. endoplasmic reticulum or mitochondria), (c) Resist denaturing, and (d) Regain their natural status on the occurrence of partial denaturation.
In other words, HSPs play important roles in assembly of protein complexes and translocation of proteins across cellular compartments, as well as in several immunological processes [28] .
In the old literature, it was believed that HSPs were 'danger-associated molecular patterns' because of being exclusively intracellular proteins only released on cellular injury or necrosis [29] . Now, it is apparent that HSPs are released from cells undergoing necrotic lysis in response to cytotoxic lymphocyte or natural killer action, or viral infections. Thus, especially members of HSP60, HSP70, and HSP90 families are capable of activating the innate immune response [30] . These members were shown to stimulate the human monocyte cell line (THP-1) to produce tumor necrosis factor (TNF-α), IL-6, and IL-8 [31] . This stimulation was mediated through different cellular receptors, either by TLR-4 (for HSP60), or CD14 (for HSP65), or by interacting with both TLR2 and 4, in a CD14-dependent manner (for HSP70) [21] . Another factor in the innate immunity role is the different abilities of HSP70s to stimulate cells of the innate immune system cells. It was found that this ability is dependent on the HSP source, whether mammalian or microbial, i.e. mycobacterial HSP70 stimulates the innate immune response, whereas some members of mammalian HSP70 may downregulate the immune response instead [32],[33] .
Adaptive immunity
Cancer studies on sarcomas indicated that HSPs could modulate the generation of adaptive immunity, and the investigators identified gp96 (a member of the HSP90 family) as the tumor rejection antigen. Immunization with gp96 elicited sarcoma-specific immunity, whereas gp96 purified from other chemically induced tumors did not elicit immunoprotection [34] . It was proposed that the immunogenicity was conferred by tumor-specific peptides associated with the HSP90 [35] , and this peptide binding is ATP/ADP dependent [36] . Interestingly, the uptake of HSP-chaperoned peptides by APCs is characterized by its availability for cross-presentation, which is the ability of exogenous antigens to enter the endogenous loading pathway of major histocompatibility complex (MHC) class I to produce CD8 + T cells [37] . The cross-presentation can occur either by the classical or the nonclassical MHC I loading, that is the vacuolar/endocytic or the cytosolic pathway [38] .
Heat-shock protein cancer vaccines
From preclinical to clinical trials (phase III), host gp96 purified from surgically removed tumors were utilized as cancer vaccines [39] . The clinical trials included tumors such as metastatic colorectal carcinoma, metastatic melanoma, non-Hodgkin lymphoma, pancreatic adenocarcinoma, and renal cell carcinoma [40] . The outcome of these clinical trials was variable, from statistically non-significant results in renal cell carcinoma, to a delay in disease progression compared with a group that received conventional chemotherapy and/or surgery in stage IV melanoma [41] . However, Colaco [42] reported that the majority of oncology animal studies used HSP70 instead of gp96.
Heat-shock proteins as infectious disease vaccines
Twenty years ago, a group of investigators found that recombinant mycobacterial HSP65 could activate murine macrophages in vivo, with intracellular inhibition of Listeria monocytogenes, but without protection against this pathogen [43] . Several studies were carried out to achieve protection in animal models using CD4 + and CD8 + T cells specific to HSP65 [44] and DNA plasmids encoding Mycobacterium leprae HSP65 [45] and HSP70 [46] . However, these studies yielded variable outcomes that were attributed to a general, rather than specific stimulation of inflammatory responses [47] . Recently, the native mycobacterial HSP16 administered with the adjuvant dioctadecyl ammonium bromide elicited protection in a TB mouse model and was capable of boosting an existing BCG vaccination [48] .
Heat-shock protein and apoptosis
In intracellular protozoons, apoptosis is considered a defense mechanism because apoptotic-infected cells are phagocytized by macrophages with parasite elimination. This phagocytosis is initiated by cellular signaling pathways, and the intracellular parasites prevent these pathways to ensure their survival in the infected cell [49] . In their study, the investigators showed that HSPs expression is directed against the parasite infection and could interfere with apoptosis by activation of the factors responsible for the regulation of anti-apoptotic molecules.
Heat-shock proteins and helminths
Schistosoma spp.
Vaccination: In the early 1990s, HSP70 was one of the antigens considered important candidates for the anti-schistosomiasis vaccine with paramyosin, triosephosphate isomerase (TPI), glutathione-S-transferase (GST), and the membrane protein Sm23 [50],[51] . Chinese investigators reported that water buffalo immunized with DNA vaccines of Schistosoma japonicum triosephosphate isomerase (SjCTPI) combined with the HSP70 plasmid and IL-12 resulted in protective immunity against schistosomiasis japonicum, with a variable reduction in all parameters (worm burden, fecal egg count, miracidial hatching rate, and hepatic egg counts) [52] . Two years later, two randomized double-blind control vaccine trials were conducted to determine the efficacy levels of the DNA vaccines encoding tetraspanin 23 kDa integral membrane protein (SjC23) or SjCTPI on their own, or when fused together with the dendritic cell targeting HSP70. They found that both vaccines incorporating HSP70 were more immunogenic and efficacious, as determined by the effect on egg burden [53] .
During 2010-2012, three Chinese articles were published on schistosomiasis vaccines. In one study, the molecular and functional characterization of S. japonicum SjMLP/HSP70 was reported. The molecule's sequence shared 77% identity with human HSP70 [mortalin-like protein (MLP)], and it was expressed widely in most developmental stages of S. japonicum (eggs, cercariae, schistosomula, and adult worms). The purified recombinant protein (rSjMLP/HSP70) was recognized by sera of infected mice. The investigators also found that the SjGST-DNA vaccine combined with SjMLP/HSP70 showed low grades of reduction (30 and 60%) of worm and egg burden in immunized mice, respectively [54] . In another study, the Chinese scientists identified in the sera of Microtus fortis, a naturally resistant vertebrate host of S. japonicum, substances with antischistosomal effects. They cloned and identified a 331 bp that was the HSP90a homologue (Mf-HSP90a). The cloned molecule showed antischistosomal effects (40-60% reduction in worm and egg burdens) both in vitro and in vivo [55] . In the third study, significant immunoprotective effects were observed when water buffalos were vaccinated with S. japonicum 23 kD membrane protein-HSP70 (SjC23-Hsp70) plus an IL-12 adjuvant. The investigators observed a reduction in all the parameters evaluated (female estrogen, fecal and hepatic egg burden) [56] .
Serodiagnosis: Four isolated HSP70 clones were evaluated for antibody activity against a panel of sera from baboons with acute and chronic schistosomiasis. The insert sizes of the four selected clones varied from 1150 to 2006 bp. It was found that antibody reactivity varied with clone length; the longest clone was the most reactive. Results suggested HSP70 immunodominance in acute as well as chronic schistosomiasis. The investigators recommended its use in the serological diagnosis of schistosomiasis mansoni [57] . Similar results were obtained on using molecules of soluble egg antigen in the diagnosis of schistosomiasis japonicum, where sera of mice with early schistosomiasis reacted with two protein molecules in soluble egg antigen: the HSP70 and 78 kDa glucose-regulated protein (Grp78/Bip) [58] . In a preliminary report, Brazilian investigators also proposed a strategy for the selection of cDNA clones that could be used in the diagnosis of schistosomiasis. Two out of five cDNA clones were identified as homologous to HSP70 [59] .
Immune response: The mechanism(s) responsible for suppression of host immune responses in chronic schistosomiasis was investigated by studying the induction of CD4 + CD25 + T cells by HSP60 of S. japonicum (SjHSP60-derived peptide; Sj MHE1). The investigators found that all mice immunized with Sj MHE1 or lymphocytes from in vitro Sj MHE1-pretreated naive mice showed an increase in CD4 + CD25 + T-cell populations as well as IL-10 and TGF-β release. The investigators suggested the possible role of TLR2 on CD4 + CD25 + induction caused by HSP60 [60] . HSP70, HSP90, and HSP97 were detected as major constituents of the proteomics profile of the excretory/secretory (E/S) products of adult S. japonicum. The investigators emphasized their central roles in immunomodulation for the host-parasite relationship [61] . In addition, Chinese scientists succeeded in cloning and constructing the recombinant plasmid of the S. japonicum HSP40 gene, which significantly initiated macrophage activation [62] .
Cancer bladder: In 2008, an Egyptian study clarified the significant association between HSP27 and HSP70 and cancer bladder caused by schistosomiasis. The investigators detected both HSPs in 60 and 68% of their patients, respectively. In addition, they found a significant correlation between HSPs expression and tumor grade and stage as well as its recurrence. They concluded that both HSPs expressions could be used as predictive biological markers for disease progression [63] .
Parasite development: Besides the morphological changes in cercaria-schistosomula transformation, cercariae have to modulate the temperature change (from external water to body temperature) and osmolarity change (from hypotonic water to saline bloodstream osmolarity). To overcome both challenges, the acetabular glands secrete HSPs as shown from the proteomic analysis of secretory products involved in host skin invasion by cercariae [64] . S. mansoni HSP70, HSP86, and HSP60 were detected in S. mansoni cercariae [65] , whereas HSP90 is a phosphoprotein detected in the different developmental stages and both sexes of S. japonicum [66] . One year later, another group of Chinese investigators emphasized the critical role of HSP in parasite development [67] . However, the heat-shock response pathway is a highly conserved and adaptive response system, and heat-shock factor 1 (Hsf1) is a major regulator of its pathway [68] . Hence, the role of gene encoding schistosomes Hsf1 in cercaria-schistosomula transformation was investigated. The results obtained confirmed the existence of Hsf1 in the cercarial stage and its expression in different schistosomal stages. The investigators identified antibodies against Hsf1 labeled on cercarial acetabular glands [69] .
Echinococcus spp.
E. granulosus : HSP70 was first reported in the serodiagnosis of hydatid cyst (HC) in 1997 in comparison with human HSP70. Both HSPs70 were assessed against patients' sera and controls. Specific antibodies' reactivity against human and parasite HSP70 was detected in 10% and 60% of the infected sera, respectively. However, 21% of the control sera reacted with E. granulosus HSP70, but the reactivity level was significantly higher in the infected patients [70] . Later, E. granulosus HSP70 was reported to stimulate the humoral and cell-mediated immune responses. Using immunoblotting, IgG, IgG4, and IgE specific to Eg2HSP70 as well as increased levels of TNF-α, interferon-g (IFN-g), IL-4, and IL-10 were detected in the sera of infected patients [71] . In addition, proteomic analysis of protoscoleces and hydatid fluid extracted from HC identified HSP20 and HSP70 [72] .
In 2010, HSP20 was identified and characterized as an immunogenic protein of E. multilocularis, and its recombinant variant showed specific reactivity to sera from infected dogs, suggesting its efficacy as a vaccine candidate [73] . One year later, also using the proteomic approach, Italian investigators succeeded in identifying HSP20 as a biological marker for the development and progression of HC. HSP20 could react with sera from HC-infected patients with different disease phases. The reaction was higher with sera from patients with active disease than in sera from patients with inactive disease [74] . Moreover, fluids from fertile HCs collected from sheep and humans as well as from infertile HCs from cattle were analyzed using tandem mass spectrometry. The investigators identified 48 proteins of parasitic origin, of which parasitic HSPs, cathepsin B and Annexin A13, were detected exclusively in infertile cysts. The investigators reported increased cellular stress and apoptosis as the major causes of their infertility [75] .
Strongyloides spp.
HSPs play a critical role in the parasite's successful establishment, its survival under stressful conditions, and its reproduction in an adverse habitat [76] . In 2009, HSB10 was characterized in Strongyloides ratti parasitic female E/S products [77] . A proteomic study of S. ratti was carried out to identify proteins found as E/S products from different developmental stages (infective larvae, parasitic females, and free-living stages). Small HSP (SrHSP10) and SrHSP60 were among the proteins abundantly observed in all the stages [78] . In the same year, another team of German investigators used mass spectrometry to identify and characterize two novel HSPs from S. ratti (SrHSP-17.1 and SrHSP-17.2) in the parasitic female E/S products. In their report, they reported the Sra-HSP-17s genomic organization, recombinant expression, purification, and antigenicity. They also showed that Sra-HSP-17s activated host cells (monocytes, macrophages, and intestinal epithelial cells), but not the lymphocytes and granulocytes. Moreover, purified rSrHSP-17s could induce IL-10 production by mononuclear cells [79] . Nouir et al. [80] from Germany worked on strongyloidiasis by analyzing the immune response to S. ratti HSP60 (SrHSP60) in infected mice. Their results showed that SrHSP60 induced Th1 response in vivo, whereas alum-precipitated srHSP60 induced Th2 response and partial protection against the challenge infection [80] . In the same year, they generated monoclonal IgM specific for srHSP60, which was used as a vaccine candidate in a mice model. Results showed a reduction in the number of the parasitic adults and migrating larvae in intestinal and disseminated strongyloidiasis [81] .
Trichinella spiralis
Levels of HSPs were used to measure stress tissue injuries caused by T. spiralis. Spanish investigators assessed HSP25, HSP60, HSP70, and HSP90 kinetics production from injured liver and muscle of infected and control mice. They found that HSP25 and HSP60 production was observed in the affected liver and infected muscles [82] . In the same year, the same investigators reported that HSP25, HSP60, and HSP70 production increased and varied in infected rat tissues (brain and spleen) according to the infection cycle, whereas a significant reduction in HSP90 was observed. The investigators did not suggest a role for HSP90 in the infection cycle [83] . However, no difference was detected in any of these HSPs in tissues of infected rats (muscle, liver, brain, and spleen) with primary trichinosis and rats reinfected 45 days after the primary infection [84] . One year later, the same investigators measured these HSPs in other infected organs (intestines, mesenteric lymph nodes, heart, and lungs) at different time points during first exposure and second reinfection. The results suggested that in reinfection, there is a sort of challenge and muscle larvae developed to adults, producing newborn larvae that were trapped in the mesenteric lymph nodes, thus increasing HSPs levels [85] . In another study, Martinez et al. [86] used three techniques to identify and purify HSPs (60, 70, and 90) from T. spiralis larvae: affinity chromatography, immunoblotting using either their monoclonal antibodies or preimmune and immune rat sera, and ELISA. Only sera from infected rats (7 days after infection) showed reactivity against the HSPs identified [86] .
A team of Egyptian scientists succeeded in obtaining HSP70 from crude somatic extracts and E/S products of adult T. spiralis, and used them to immunize mice against challenge infection. The resulting strong immunogenic reactions of HSP70 were evaluated by parasitological measures (worm and muscle larval burden) and increased total g-globulins [87] . The first molecular cloning and characterization of HSP70 from T. spiralis was performed in China. The recombinant TsHSP70 formulated with Freund's adjuvant proved to be a possible candidate vaccine against trichinosis as shown in the strong humoral immune responses in mice challenged with T. spiralis larvae [88] . Two years later, screening of cDNA libraries from T. spiralis adult worms against sera from infected pigs produced several positive clones. Of these clones, 14 showed sequence identity with HSP70, suggesting its major role in host-parasite interactions [89] .
Filarial nematodes
In lymphatic filariasis, there is impaired monocyte functions indicated by reduced production of chemokines (e.g. IL-8, eotaxin, and Exodus II), and cytokines (e.g. IL-1α, IL-12p40, IL-12p70, IFN-g, and IL-10) [90] . A group of American scientists assessed gene expression patterns of monocytes from infected patients and healthy individuals. They recorded differences in expression among the genes involved in apoptosis and cell adhesion. They further compared these gene expressions to before and after treatment (single dose of ivermectin-albendazole), and the most detected alterations were found in the expression of HSP60, HSP70, HSP90, and HSP105 genes. The investigators concluded that treatment resulted in the clearance of the parasite's antigens that retained monocyte-suppressive functions, releasing chemokines and cytokines as well as increasing HSPs expression [91] . However, it was reported that recombinant Wolbachia (microfilarial symbiotic bacteria) HSP60 (rwmHSP60) regulates immune activation in lymphatic filariasis by inhibition of secretion of proinflammatory cytokines from activated T cells through TLR signaling. Decreased CD69 with elevated CD127 and CD62L as well as IL-10 and TGF-β in infected patients confirmed reduced T-cell activation because of HSP60 stimulation [92] . One year later, Indian investigators reported that rwmHSP60 also induced proinflammatory cytokine production (IL-1β, IL-6, and TNF-α), reduced phagocytic function of monocyte/macrophage, and promoted apoptosis of human monocytes [93] .
Wuchereria bancrofti and Brugia spp.: Brugia spp. (malayi and pahangi) HSP70 was first discovered in 1987, and was described as a protein expressed in both the insect and host parasitic stages. The investigators reported that it played no role in host immune response [94] . At the same time, they used the C-terminal portion of Brugia spp. HSP70 (Bpa-26) as an indicator for chronic Brugian filariasis. The investigators found significantly higher IgG3 levels in chronic patients compared with microfilaremic and asymptomatic amicrofilaremic patients [95] . The recombinant protein of Brugia malayi Wolbachia HSP60 was also reported to generate antibody response (total IgG and Ig1) in all individuals from an area endemic for W. bancrofti [96] . In 2010, HSP70 identified from the bovine filarial parasite Setaria cervi showed 98% identity with that of W. bancrofti and only 28% with human HSP70 [97] .
The specific inhibitor of B. pahangi HSP90 such as geldanamycin was reported to kill adult worms and microfilariae in vitro. However, it proved to have severe hepatotoxic hazards besides being metabolically and chemically unstable [98] . One year later, the same investigators developed a fluorescence polarization assay that proved sensitive and specific in identifying new compounds that bind to the HSP90 pocket and could be used as an HSP90 specific inhibitor. The investigators validated their assay and its adaptation to screen HSP90 inhibitors against other parasites [99] . In a study carried out by American and Indian investigators, the vaccine potential of B. malayi HSP12.6 (BmHSP12.6) was evaluated in infected mice. The results showed that vaccinated mice had ~72% protection against the challenge infection. The investigators suggested that BmHSP12.6 had host immunomodulatory functions that might interfere with microfilaria establishment in the host. Moreover, there was a significant increase in IgG1 and IgG2a antibody responses. They concluded that BmHSP12.6 could be developed into a vaccine candidate against B. malayi [100] . One year later, BmHSP12.6 was used as a partner in a trivalent fusion vaccine, and conferred significant prophylactic protection (95%) against challenge infection [101] . Recently, B. malayi HSP12.6 (which is an a crystalline domain and c-terminal extension) was included in a vaccine with a B. malayi abundant larval transcript (rBmALT-2) and a B. malayi tetraspanin large extracellular loop (rBmTSP LEL) to form rBmHATαc [102] . It conferred 68% protection in mice with challenge infection that increased to more than 95% protection when AL007 (alum from IDRI) or AL019 (alum plus TLR-4 agonist from IDRI) was used as an adjuvant [103] . More recently, German scientists investigated the role of B. malayi HSP70 in immunizing BALB/c mice against challenge infection with Litomosoides sigmodontis (rodent filarial worm). The results indicated reduced numbers of pleural cavity larvae and circulating microfilariae as well as increased production of type I and II cytokines. The investigators concluded its potential role as a vaccine candidate [104] .
Onchocerca volvulus: HSP70 was first reported in 1989 when investigators recognized three cloned filarial immunogenic antigens that reacted with sera from infected patients. One of these immunogenic antigens recognized by amicrofilaremic patients is an O. volvulus protein homologous to the 70 kDa HSP of the African frog Xenopus laevis [105] . Later, chaperonin 60 (CPN60) or HSP60 of O. volvulus (OvCPN60) was reported in 2000 and the investigators detected it in all life-cycle stages. Antibodies raised against OvCPN60 were found in ~90% of infected patients in West Africa [106] . In 2008, a group of German scientists reported that killing Wolbachia bacteria in O. volvulus-infected patients resulted in an increase in HSP60 production from filarial mitochondria. To differentiate between HSP60 from Wolbachia and that of filarial mitochondria, they used antiserum raised against HSP60 to specifically bind to the protein and stain it in both Wolbachia and mitochondria. They attributed this increased expression to homeostasis disruption of endosymbiosis [107] .
Other helminthes
Taenia solium: A small HSP, Tsol-sfISP35.6, was isolated by antibody screening of a T. solium cDNA library, followed by cloning. Using western blot analysis and ELISA, the clone reacted with 80% of sera of infected pigs, 84% of patients with active neurocysticercosis, and 71% of patients with inactive diseases [108] .
Echinostoma spp.: Using a proteomic approach, Spanish investigators identified protein spots corresponding to 10 proteins including HSP70 in E. friedi E/S products. The stress protein showed variable expression patterns in chronic and acute infections in experimentally infected rodents, suggesting its role in parasite survival [109] . HSP70 was also identified in E. caproni E/S products. Another group of Spanish investigators recognized an early IgM response generated by HSP70, suggesting its importance in parasite establishment because of its early exposure to the host immune response. Moreover, IgA response was mediated by HSP70, which was attributed to its close contact with the host mucosal surface [110] .
Paragonimus westermani: In 2007, the P. westermani egg antigen gene was cloned and evaluated as an antigen for the serological diagnosis of paragonimiasis. Using ELISA against sera from patients infected with different parasites (including patients with paragonimiasis) and negative controls, its sensitivity and specificity were 90.2 and 100%, respectively. Moreover, its sequence was 40, 38, and 35% identical to HSPs from S. japonicum, S. mansoni, and T. saginata, respectively [111] .
Fasciola spp.: In 2008, F. hepatica and F. gigantica HSP70s were cloned and characterized. Using Western blot analysis, Australian investigators found 98-99% identity in their protein sequences. However, the 70 kD protein was not detected in the parasite E/S products. Analysis of HSP70 expression in fascioliasisaffecting sheep (not cattle), showed higher HSP70 expression in F. gigantica than F. hepatica. This result highlighted the importance of HSP70 as a biochemical marker of F. gigantica stress response [112] .
Clonorchis sinensis: In clonorchiasis, HSP27, HSP90 expressions as well as levels of inflammatory cytokines (IL-1β, IL-2, TGF-β2, and IFN-α1) were increased in infected hamsters compared to control group. This significant increase was correlated with the severity of bile duct hyperplasia, liver fibrosis, and hepatic necrosis [113] .
HSPs play important roles in the correction of cell protein functions under stress conditions and share in several immunological processes of innate and adaptive immunity. Their role in the vaccination era was reported against cancer (such as cancer colon, renal cell carcinoma, and pancreatic adenocarcinoma) as well as against microbial and parasitic diseases. HSPs expression was also found to be directed against apoptosis, especially in intracellular protozoon.Several studies were carried out using HSP70 as a vaccine candidate in schistosomiasis japonicum. In addition, few studies suggested the use of HSP70 in the serological diagnosis of schistosomiasis mansoni.HSP70 and HSP90 were detected as major constituents of the proteomics profile of the E/S products of adult S. japonicum, and they were impacted in host-parasite relationship immunomodulation. However, HSP70, HSP86, and HSP60 were detected in the secretory products of S. mansoni cercariae, and they play a critical role in parasite development.In hydatid disease, E. granulosus HSP70 was observed as an immunogenic protein stimulating both the humoral and the cell-mediated immune responses. Use of E. multilocularis HSP20 was suggested as a vaccine candidate.In strongyloidiasis, small HSP (HSP10) and HSP60 were observed in all the stages of S. ratti. Other sHSPs (HSP17) were detected in the parasitic female E/S products, which activated most host cells and induced IL-10. However, HSP70 was found to induce Th1 response in vivo, suggesting its use in protection against challenge infection.Several HSPs were reported to be increased during stress tissue injuries in trichinosis such as HSP25, HSP60, and HSP70. There is some contradiction on HSP90 production from the injured tissues. However, HSPs 60, 70, and 90 from T. spiralis larvae showed reactivity against sera from T. spiralis-infected rats.T. spiralis HSP70 produced strong immunogenic reactions when used to immunize mice against challenge infection.HSP70 of both W. bancrofti and Brugia spp. were recognized as immunogenic proteins capable of significant production of IgG3 levels. In addition, B. malayi HSP70 was used to immunize mice and showed its potential role as a vaccine candidate. However, B. malayi HSP12.6 was used as a partner in a trivalent fusion vaccine (rBmHATαc), which conferred excellent protection in mice if used with AL007 or AL019 as adjuvants.Few other studies have focused on HSPs in other diseases caused by helminths, for example O. volvulus, in which HSP60 was detected in all life-cycle stages. A clone of HSP 35.6 in T. solium reacted with the majority of sera from patients with active neurocysticercosis. HSP70 was detected in different Echinostoma spp., and it was found that IgA response was mediated by HSP70. In fascioliasis, HSP70 showed high potentiality as a biochemical marker in infection with F. gigantica. In clonorchiasis, HSP27 and HSP90 expressions were elevated with increased production of inflammatory cytokines.
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My Weblog: Let it snow!
Yes, I'm late posting this blog. No excuse, except that I've been so focused on the weather that it only just dawned on me that today was Monday.
Full confession here: I am a snow wimp. I am one of those people who makes snowstorms in the Washington Metropolitan area so amusing to people from many other parts of the country. Not sure which is worse: the Midwesterners and New Englanders laughing at how we go crazy at what they consider minuscule amounts of snow or the Floridians and Californians who feel obliged to tell us how balmy their weather is while we're shoveling white stuff. Sometimes I find it better to stay off of social media during a snowstorm. Better for my peace of mind.
And this snowstorm was made slightly more stressful than usual by the fact that for a variety of complicated reasons, I ended up being the person available to take my 15-year-old nephews to a four o'clock basketball game out in the next county. My natural reaction to news that a snowstorm is approaching is to make sure I have a supply of comfort food—forget the bread and milk; we're talking Diet Coke, some form of potato substance, and whichever species of lettuce is currently deemed low risk—and hunker down. Long before the first flake falls, I want to be sitting at the computer or in my recliner, facing no challenges more daunting than whether to read my book, watch Netflix, or start cooking dinner.
Didn't quite happen that way. I admit, I was a little anxious as I headed out, and the boys, who seem to have inherited their material grandfather's love of teasing, took care to point out every place where the snow was sticking. And by the time the game was over, it was sticking all over the place. But the roads weren't actually that bad, so in spite of being a snow wimp, on the way home I agreed to a detour to pick up Chinese food. And stayed to eat it with them before scampering home to cocoon.
Am I becoming less of a snow wimp? Probably not. But for the nephews, I'll brave a little snow.
Actually, rather a lot of snow. My neighbor who has been out cross-country skiing in the woods behind our houses estimates ten inches, maybe more. I don't see any reason to argue with him. Statisticians say this weekend's storm was the ninth largest January snow on record, and the seventh biggest two-day storm in the past 30 years. For those of us able to stay in for the duration, it was a surprisingly undramatic storm, depositing those 10-12 inches slowly but inexorably over a full 36 hours. And without causing widespread power outages. A civilized storm.
Still, I was glad when my snow guy showed up, a cheerful Spanish guy with whom I have no contract, but who can be relied on to show up after every snowfall and clear my driveway and walk beautifully for what I consider a reasonable price. And even more glad when I was able to get out today to run a few errands.
And I was ready for some snow, in part because I'm now working on Owl Be Home for Christmas—another Meg Christmas book. In which there will be snow. Vast amounts of it. It's going to be one for the record books. (In the DC area, the record is 28", set in 1922. In Richmond, almost 22" in 1940. I think we'll plan for about three feet. Brace yourselves, Caerphillians!) So having a reasonably well-behaved if highly productive snow storm show up when I'm in the middle of writing about my characters being snowbound seems almost useful.
Here's hoping that if you were in the path of this recent storm, you're inside safe and warm with clean sidewalks and driveways and a plentiful supply of your preferred storm supplies!
So, it sounds like you want to hear about the weather here in CA. We actually got 2 inches of rain yesterday, and it didn't make it to 50. Today will be more of the same. No snow, but as wintry as we get.
Oddly enough, I think I would miss snow if I never had any. That could be the mellow, survived-another-snowstorm me talking.


The other day at a play date the kids discovered Floam and have been wanting to play with it at home. After searching countless stores and leaving empty handed we decided to attempt to make some. Making your own play supplies with your kids is a fantastic way to sneak some learning into their play. We had a blast experimenting with how to make our own Floam!
If you have never played with Floam before, it is definitely unique. We made our homemade floam in a similar manner to slime and our DIY Kinetic Sand.
Just like the Kinetic Sand, the consistency of your floam is going to depend on much of the borax mixture you add in. For a firmer Floam you will want to use more borax. I would suggest starting out light and slowly add it in until you find a consistency that you like. This is also an awesome chance to work with your kids on hypothesizes!
Floam can be stored in an air tight container for endless play, or kids can leave their creations out to dry. We have yet to let ours dry out intentionally, but we have had some dry out on us over night. I am thinking this would be a super fun material for a sculpture project! Also Floam left in a ziplock bag makes a great "stress relief" ball!
For your Floam filler, polystyrene beads can be found in most craft stores for an affordable price. If you cannot find them they are available for purchase on Amazon or you could try breaking up any Styrofoam you have on hand.
In a bowl combine 2 teaspoons of borax with 1/2 cup of very warm water.
Mix until the borax is dissolved.
In a separate bowl combine 1/4 cup of Elmer's glue with 1/4 cup of water. Add tempera powder if color is desired.
Once the ingredients of both bowls are dissolved pour everything into a ziplock bag (glue mixture, borax mixture, and polystyrene beads).
Seal the bag and shake until everything is combined.
This unique Sensory Material is also super easy to clean up! Hope your family enjoys this as much as ours has!!


When my T430s arrived, OpenBSD didn't yet support Sandy/Ivy Bridge graphics, so I stuck with Windows and OpenBSD in VMWare. Things change and now I want to run OpenBSD natively. I'm using TrueCrypt on the whole drive and trying to resize that while introducing another boot loader seemed a risky proposition, so I cheated a little by taking advantage of my laptop's mSATA port and installed a 64GB Crucial m4 mSATA SSD, a trivial upgrade.
Plug in an external hard drive I keep around for booting OpenBSD and boot bsd.rd. In keeping with protecting all the data on this laptop, I encrypted the whole drive as well. By the time I got to the disk setup part of the installer, I was up to sd3 (Samsung SSD, m4 SSD, USB disk, softraid). A few years ago I would have marveled at installing OpenBSD on a laptop with 4 "SCSI" disks.
OpenBSD runs well, as expected, except for the USB3 ports which are completely dead. In order to get turbo boost modes out of the CPU, I have to goose hw.setperf after boot. My theory is that we boot in the P1 state, but the kernel assumes we are in P0 because the current frequency matches the CPU's designated operating frequency. (I'm supposed to know more about this, but have kind of forgotten the particulars of setperf and cpu scaling.) Suspend and resume appear reliable.
X works on the native panel and with an external monitor, although it requires fiddling with xrandr. I unfortunately discovered that xrandr likes to reset the dpi setting, and X's (or X apps') device independent scaling is total balls, so the first thing to do after getting all the screens set up is running xrandr --dpi 96 again. By default, two finger scrolling works, but tap to click doesn't. xinput --set-prop 7 "Synaptics Tap Action" 0 0 0 0 1 3 is the magic command to fix that.
Back on the Windows side, some fiddling with VMWare lets me boot the disk as a VM, enabling data transfer between disks. Be careful not to suspend and resume the VMWare image, though. softraid prints a roaming disk warning about the device changing, but copes with the change. Using DUIDs in fstab makes the whole thing transparent.
Performance is good. I can get about 250MB/s reading from the raw Samsung 830 SSD and 170MB/s from the m4. I think the 128GB or 256GB models may offer better performance having more NAND dies, and 128GB is certainly at a better point on the price/GB curve, but 64GB was more than sufficient for my OpenBSD storage needs. softraid cuts the read performance from the encrypted drive down to 120MB/s, so I think there's some optimization work that can be done there, but it still feels fast.
^ This is where I think switching between 2601 and 2600 makes all the difference with turbo boost.


The idea of creating a business vision is not new. Many business owners start their business with a vision and some even create a strategy to achieve it.
However, many business owners fail achieve their vision and that's usually because they fail to take the actions necessary to make it happen.
Follow these 3 steps to create a business vision that actually motivates you to take action!
Firstly, a vision is very simply a picture in the imagination and it's usually a long-term view of the future. In a business context, it is an image of how the business owner wants the business to look in the future and so it provides the direction of the business. It includes the core values that drive the ethos and culture of the business and it forms the basis of the strategy and all activity going forward, providing a framework within which all decisions are made.
An effective business vision is one that inspires and motivates the people in the business to take action! If it doesn't do that, then it is of little use.
Your Intent is about the 'what'. It's about desire. In other words, it's about 'what' you 'want'.
It's about knowing what you want your business to be like. It's about defining what you want to achieve specifically and it's about clarifying the kind of work you want to be doing and with whom. It's also about deciding where you want to work from, both in terms of environment and location.
Purpose is about the 'why'. It's about your reason. In other words the 'reason' you are in business.
Your 'why' stems from your personal values (the things that are important to you) and because of that, it is your deepest driving force. It's about what your intent means to you and specifically what achieving it will enable for you (and for those you care about).
Once you have defined your intent and purpose, you have the basis of a vision. However, you want your vision to exist as reality at some point in the future, which means that it needs to inspire and motivate you to take the kind of action that will make it so.
The key to this is making it 'tangible'. What I mean by that is creating a tangible version of your vision, so that it exists in some form, outside of your head. Think of creating a tangible vision as literally creating your future history. In other words, creating the outcome as if it exists right now. The reason for this is that when your vision exists in some form other than in your imagination, your beliefs, thoughts, feelings and importantly, your actions become aligned with it, and you are more likely to make it happen!


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Newsom's recall fundraising edge becomes a chasm with union, tribal, tech money
Dustin Gardiner, Nami Sumida
June 17, 2021 Updated: June 22, 2021 5:50 p.m.
California Governor Gavin Newsom waves after a news conference at Universal Studios in Universal City, Calif. on June 15, 2021.
Ringo H.W. Chiu/Associated Press
SACRAMENTO — There was always little doubt that Gov. Gavin Newsom would have the financial edge in California's recall election, but the fundraising gap between the governor and his main GOP challengers has become a quickly growing chasm.
Newsom, a Democrat, has raised nearly $16.5 million to defend himself in the recall contest expected this fall, with much of the money pouring in over the past two weeks. He has also raised $9.3 million for his 2022 re-election campaign.
In other words, the governor has pulled in about twice as much as his entire field of Republican challengers combined. The next closest candidate, San Diego businessman John Cox, has raised $7.7 million, most of it from his own pocket.
The governor has been boosted by a cash infusion from mega donors, including labor unions, the Democratic Party, big tech billionaires, tribal communities and real-estate developers — a broad coalition that melds both traditional progressive donors and business interests.
Several factors have helped boost Newsom's advantage: For starters, he can raise unlimited funds to fight a recall; state contribution limits don't apply to recalled officeholders. Donors to his Republican challengers, on the other hand, are limited to the normal $32,400 maximum per campaign.
Democrats propose $215 million to pay for, speed up Newsom recall election
When will California vote on Newsom's recall? Here's what the person in charge of setting the date says
Newsom has also benefited as California emerges from the coronavirus pandemic with a booming economy and an unexpected $75.7 billion budget surplus. His campaign released its first television ad Thursday, declaring the state is "roaring back" and highlighting a plan to send many households $1,100 tax rebates.
David McCuan, a political scientist at Sonoma State University, said timing has clearly been on Newsom's side. He said the governor has seized on that good fortune and gradually built a fundraising "tsunami" to drown out opponents on the airwaves.
"That's a really hard background to raise money if you're any Republican," McCuan said of California's pandemic rebound. "The dollars are a reflection of the stock price of the candidate, and a forward looking average."
Newsom's largest checks include $3 million from Netflix CEO Reed Hastings, $1.5 million from the California Association of Realtors and $1.32 million from the state Democratic Party, including some in-kind services such as digital support.
However, Newsom's opponents say the governor's fundraising lead isn't necessarily an indicator of how voters feel. Several accused the governor of using his office to benefit large donors like state-employee labor unions.
Cox, whom Newsom defeated in the 2018 governor's race, said Newsom's advantage is to be expected when "politicians do favors for people." He said he expects that voters frustrated with the state's myriad problems, including power shortages in the summer and the homelessness crisis, feel differently than wealthy donors.
"So many things are mismanaged in this state that I won't need but a tiny fraction of what he's able to get from his cronies and favor seekers," he said of Newsom.
Cox, a multimillionaire and tax attorney, said he plans to spend even more of his own money to try to defeat the governor, though he declined to say how much.
Newsom's next closest competitor, former San Diego Mayor Kevin Faulconer, has about $2.4 million. Despite less fundraising, a spokesperson said Faulconer has a growing base of supporters and is "confident that our campaign will have the resources and support we need to defeat Gavin Newsom's big money political machine."
Faulconer's biggest contributors include a host of Southern California real-estate developers. Among them were three members of the Marcil family, who own Palos Verdes Investments, a company that redevelops apartment complexes.
Combined, the Marcil family gave over $194,000 to Faulconer's campaign committees to run for governor in 2021 and 2022. Candidates can raise for both committees simultaneously.
About Our Newsroom
Our politics team covers California government from Sacramento and national politics from the Bay Area and Washington, D.C. The guiding principle in choosing which stories to cover is: How does political and government news affect the Bay Area and California?
Read more about how The Chronicle covers politics and what we do to ensure fairness in our reporting
For more on the recall election, see our special report.
The two other prominent Republicans in the race, reality TV star Caitlyn Jenner and former U.S. Rep. Doug Ose, have lagged far behind in reported fundraising. Jenner has raised about $295,000 while Ose has reported about $157,000.
Jenner's biggest donors include a handful of wealthy Republicans from outside the state. For example, Arizona billionaire Bob Parsons, who founded the internet domain company GoDaddy, and his wife, Renee Parsons, each gave $32,400.
Nathan Click, a spokesperson for Newsom's campaign, dismissed attacks from his opponents, saying, "The governor is in a position of strength. We're not taking anything for granted."
McCuan, the political scientist, said it's clear that to compete against Newsom's fundraising operation, GOP challengers will need to appeal to conservatives outside of California. So far, they've all struggled to lure big, out-of-state donors, despite a flurry of Fox News and conservative media segments focused on the recall.
"The way that Republicans can raise money is to go outside the state and nationalize and demonize (Newsom)," McCuan said. "That's what it will take."
Dustin Gardiner and Nami Sumida are San Francisco Chronicle staff writers. Email: dustin.gardiner@sfchronicle.com, nami.sumida@sfchronicle.com Twitter: @dustingardiner, @namisumida
Dustin Gardiner
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Dustin Gardiner is a state Capitol reporter for The San Francisco Chronicle. He joined The Chronicle in 2019, after nearly a decade with The Arizona Republic, where he covered state and city politics. Dustin won several awards for his reporting in Arizona, including the 2019 John Kolbe Politics Reporting award, and the 2017 Story of the Year award from the Arizona Newspapers Association. Outside of work, he enjoys hiking, camping, reading fiction and playing Settlers of Catan. He's a member of NLGJA, the association of LGBTQ journalists.
Nami Sumida
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Nami Sumida is a data visualization developer at The San Francisco Chronicle. Before joining in 2021, she created data-driven graphics for Industry Dive, a business journalism company covering 20+ industries. She also has experience conducting research on journalism and the news media at the Pew Research Center.
Read more about the data team and their work.

redpajama_set_name: RedPajamaCommonCrawl


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text:
Gauging Groundwater
The largest assessment of global groundwater wells finds many are at risk of drying up
Global groundwater wells at risk of running dry
Scott Jasechko.jpeg
Scott Jasechko
"Jack and Jill went up the hill to fetch a pail of water." It's a silly rhyme, but one that highlights a simple fact: Humans have long relied on wells — such as the one on the hill visited by Jack and Jill — for their primary drinking water supply.
Although the number of people who draw their water by pail is declining as pumps become ever more widespread, groundwater wells still supply drinking water to more than half of the world's population and sustain over 40% of irrigated agriculture. But this vital resource underfoot often gets overlooked.
UC Santa Barbara assistant professors Debra Perrone and Scott Jasechko have compiled the most comprehensive assay of groundwater wells to date, spanning 40 countries that collectively account for half of all global groundwater pumping. Their study, the cover story of the April 23 edition of the journal Science, offers an unprecedented account of the world's groundwater resources.
"We analyzed construction records for tens of millions of groundwater wells around the world," said Jasechko, of the university's Bren School of Environmental Science & Management, "and the big take home is that many wells are at risk of running dry." Up to one in five wells in their survey, to be exact.
"These groundwater wells are used to withdraw water for household use or irrigation," added Perrone, a faculty member in the Environmental Studies Program. "Our study is the first time that groundwater wells and water level observations have been stitched together like this, providing local insights at the global scale."
Reaching these conclusions required the better part of six years' work by Perrone and Jasechko. The researchers culled their data from more than 100 unique databases around the world, sometimes in different formats and languages.
The dataset contains roughly 39 million wells in 40 countries or territories. Blue points mark shallower wells and red points mark deeper wells.
Photo Credit: PERRONE AND JASECHKO
The first step involved compiling roughly 39 million records of functional groundwater wells that included depth, purpose, location and construction date. Rather than delegate processing the data to an algorithm or AI, the researchers carefully went through tidying it up manually. "We wanted to be sure we understood the limitations and nuances for each database we analyzed," Jasechko said.
With the data sorted, the authors compared the depths of local water tables against well depths. They found that 6 to 20% of the wells in their global sample were at risk of running dry if water levels continue to decline by just a few meters. They sought to find trends in the depths of new wells compared to existing wells in a given area from 1950 through 2015. While newer wells tended to be deeper than older ones, the trend was not ubiquitous, they said. Functional wells were not getting substantially deeper in many of the areas they surveyed.
Perrone stands next to a groundwater well used to monitor natural conditions at the Jack and Laura Dangermond Preserve.
Photo Credit: COURTESY IMAGE
The researchers then pulled data from roughly 1.1 million monitoring wells, which provide measurements of groundwater levels and conditions. These records supplied not only an account of where the water table lay, but also how much it fluctuated over the course of the year. Seasonal variation, they found, was typically within one meter.
By comparing depth trends to measurements from nearby monitoring wells, Perrone and Jasechko could also deduce how close functional wells were to drying out. They found that in many areas groundwater levels are declining, yet new wells are not being drilled deeper to keep pace with the falling water table. Since wells aren't getting substantially deeper in many areas, newer wells are at least as vulnerable as older wells should groundwater levels continue to decline.
"In some places, groundwater levels are close to the bottom of wells," Perrone stated. "If groundwater levels continue to decline in these places, wells will go dry, leaving people without access to water."
The two researchers also availed themselves of data from NASA's GRACE mission. The pair of GRACE satellites detect small differences in the Earth's gravitational field as they orbit the planet. This provided Perrone and Jasechko with information about groundwater reserves for regions where data from monitoring wells is limited.
"We only have groundwater level monitoring data for a few dozen countries," Jasechko said, "but GRACE data allows us to explore changes in water storage around the globe." While the resolution was coarse, the additional information confirmed the disconnect between declining groundwater and the construction of deeper wells.
There are many reasons not to drill deeper. It's much more expensive in terms of drilling and operational costs. The quality of groundwater can also decrease at depth, often becoming more saline toward the bottom of an aquifer.
"Drilling wells is expensive," Jasechko remarked. "Even if fresh water exists deep underground, not every individual or household has the capital to drill a new well to access it, which raises concerns about equity when wells run dry."
The pair hope to expand the coverage of their dataset, especially to countries like China, Iran and Pakistan, three major users of groundwater that the researchers could not obtain records from. They also plan to investigate the extent of groundwater decline in future research, looking for information on how quickly water tables are dropping and where the trend is accelerating.
Groundwater can provide a lifeline in even the most arid regions.
Perrone and Jasechko have made a name for themselves in the field of groundwater research by approaching the topic with enormous datasets like this one. The global study comes on the heels of another paper published in Nature on groundwater resources across the United States. There, they revealed the extent to which rivers across the country are losing flow to aquifers, in part due to pumping.
Together, the findings paint a broad picture not just of wells in danger, but of dwindling groundwater resources as a whole. While pumping may have the most direct impact on aquifers, the authors note, human activity has many indirect effects on groundwater quality and levels. Industrial activity, water diversion and even pavement and runoff affect the water beneath our feet. Climate change may exert the greatest influence over the long term, as it alters the distribution and intensity of precipitation and evapotranspiration from plants and soils worldwide.
"These findings highlight the importance of groundwater resources and groundwater management to the global community," Perrone said. "Putting all the data together reveals that declining groundwater levels threaten wells in many places across the world."
Groundwater sustainability is a complex issue with a lot of considerations and tradeoffs, the authors acknowledged. Deciding on a course of action won't be easy, but compiling and analyzing large datasets like this are a critical first-step in tackling the challenge. "With these data, we can make more informed management decisions to help us use groundwater sustainably," Perrone said.
Debra Perrone
A System for Success
Biodiversity Imperiled
Losing Rivers
A Well-Rooted Study
The Vital Resource Beneath Our Feet
Drilling Deeper
An Opaleye-opening Result
Managing Water Resources in a Low-to-No-Snow Future
Hot Town, Summer in the City
Feeling the Squeeze
Shrinking Groundwater


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RealEstateReview.com.au Australian real estate and property news, reviews and updates
Australian Property Updates
Property Information & Real Estate Updates
Sellers & Buyers
Home Buyers Info
Home Buyers Mistakes
Home Sellers Info
Home Seller's Mistakes
Buying Before Selling
Home Renovators
Renovation Steps
Renovate Or Move
Make Money in Renovation
Business & Franchise
Agents required to disclose the history of the property
Agents are required to disclose the history of the property, even if a murder took place in a house they are selling. The Australian Competition and Consumer Commission this month gained power to fine agents up to $1.1 million for misleading their clients. Under the new laws, if properties such as the Glen Iris townhouse where Margaret Wales-King and her husband Paul King were bludgeoned to death or the Mornington home where John Sharpe killed his wife Anna Kemp and daughter Gracie with a speargun were to come on the market, potential buyers would have to be told that murder had occurred in the house.
Agents failing to disclose a property's past to a buyer could face a million-dollar fine. Concealing knowledge of noisy neighbours or barking dogs will also attract penalties. The changes came into effect on July 1 when the ACCC merged state fair trading laws into a federal consumer protection law. Real Estate Institute of Victoria chief executive Enzo Raimondo said there had previously been no penalty for failing to disclose a murder house when asked.
He said tightened disclosure laws would give consumers more confidence when buying homes. "It's about being fair and honest to everybody and if the potential purchaser has asked a direct question the agent should say: 'Yes that is the property where the murder happened', if the agent knows about it," Mr Raimondo said.
There had never been a case in Victoria where an agent had failed to disclose a murder when asked, he said. In 2004, two Sydney agents were fined $20,000 after concealing from a buyer the fact that a house had been the scene of a triple murder committed by Sef Gonzales.
Source: news.com.au
Ethan Billson
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The information on RealEstateReview.com.au has been prepared without taking into account your financial situations. This is general info.


Lyme disease is an affection caused by a spirochete infection called Borrelia Burgdorferi which may harbor a varied and misleading clinical symptomatology. The serology tests commonly used for diagnosis show a wide sensitivity varying from 34% to 70,5%, leaving many infected patients with false negative tests. Alternative techniques such as polymerase chain reaction (PCR) could be helpful but not conclusive enough. Using biofilm busters, such as stevia and serratiopeptidase, could lead to bacterial blood release, thus increasing the spirochete load, making PCR test more sensitive, thus improving the patient 's diagnosis and management.


I'm so excited about the 2018-2019 school year; it's off to a great start! Thanks for visiting my website! Please click on the heading to the right that best represents you (teacher, parent, student) and you will find an array of information suited to you. This is updated often, so continue to check back throughout the school year for additional information.


We recommend a second person to help transcribe measurements while you measure for your 1/4 Inch Skins.
Skins are the ¼ inch plywood that you put on top of your boxes. We recommend to measure the under cabinet first; this is the section below the upper cabinets. There may be a section that divides the under panels so you will need to take separate measurements for each side. Try to be as accurate as possible when making these measurements.
After measuring the upper cabinets, move onto the left side of the kitchen. If you have a large panel with a microwave and oven, go ahead and measure all the way to the crown. Repeat these steps for the other side of the cabinet.
If you have a backsplash or countertop you will need to add a quarter inch to make sure your measurements aren't too small.
The first thing you need to do is idenify the different widths for your face frame material. Typically, the widest section is the Toekick. This is usually the area that sits behind your face that comes up to the bottom of the cabinet from the floor. The best way to double check to see if the toekick is the widest area is to open all the doors and drawers and visually look at the width of your styles and rails.
It's a good practice to order at least a ¼ inch wider skin than what it measures. All the material comes in 8-foot length. When you're making your count. Count them in 8-foot sections. After locating the widest section move your way down to the rest of the cabinets so your recording from widest to narrowest.
4. How Many Sticks of Each Width Do I Need?
Go back to your recorded widths and look before measuring. Keep in mind everything comes in eight-foot sticks so that's what you need to calculate. It's good practice to add two extra sticks just to the count for any cutoff waste.
Remember the Carpenter's Rule: "Measure twice, cut once." Start by counting your verticals first then move onto your horizontals.
Order sticks in even numbers so they stack evenly without shipping damage.
Double-check your measurements by repeating steps 1 through 4. Remember to always measure width first then height. If you're not comfortable with your measurements go through them again. It's better to have extra material than not enough.


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Home / Volvo
By Armen Hareyan G+ Sep 25 2020 - 8:22pm
2020 Volvo XC40 Inscription: Award-Winning Technology and a Cool Crystal Gear Lever
The new 2020 Volvo XC40 is Volvo's third and smallest SUV, but the award-winning technologies from the largest XC90 and XC60 series are standard in the 2020 Volvo XC40. These are innovations, which Volvo says are rarely offered among competitors.
This week Torque News test-drove the 2020 Volvo XC40T5 AWD Inscription, which is the top trim of the series, offered with Momentum and R-Design trims. The top characteriestics of the Inscription trim are the exclusive design, luxury by Sweden, the premium touch and the right stance. However, the Volvo XC40 is also offered as a fully electric SUV (XC40 Recharge) with more than 200 miles of range and 78-kwh battery pack.
XC40's Inscription Trim
Let's talk about some of the characteristics of the 2020 Volvo XC40's Inscription trim. Once you seat in the car, the first thing that stands out is the crystal gear lever. It really creates a contemporary sense of craftsmanship in the cabin, providing nice reflection of light during night driving. Another design touch that makes the Inscription trim to stand out is the chrome trim of the front grille and side windows. The roof rails are made of bright aluminum, and all of these distinctive features give the vehicle an exclusive design. Other distinctive characteristics of the Inscription XC40 are the rich leather upholstery in the interior and the striking 18, 19 or 20-in alloy wheels.
Pricing and Fuel Economy
The 2020 Volvo XC40 T5 AWD Inscription starts at $35,700, but can go up to $47,395 when you add all options. This SUV gets 25 MPG combined, while the City and highways are respectively 22/30 MPG.
Volvo is known for its large cars and especially its station wagons, which are among the safest vehicles in the world. The Swedish manufacturer has succeeded in carrying on this tradition with a new range of vehicles by introducing its first SUV on the market in 1997. What's interesting, is that the Volvo XC40 fits perfectly with the image we have of a modern SUV.
Despite its small size the Volvo XC40 remains quite powerfull under the hood. From the entry-level version, we are treated with a small 2.0-liter 4-cylinder turbocharged engine, developing 188 horsepower and 221 lb-ft of torque. This mechanic is called T4 if you are familiar with Volvo terms. The second option is also a turbocharged 2.0-liter 4-cylinder engine, but the output of the latter increases to 244 horsepower and 258 lb-ft of torque. Despite its small size, the Volvo XC40 remains quite heavy with a total weight of over 1,600 kg. In other words, that powerful engine is not too much.
In either case, the 2020 Volvo XC40 benefits from an 8-speed automatic transmission and all-wheel drive, which allows it to deliver a sporty drive while maintaining excellent fuel consumption.
The Volvo XC40 is a true luxury vehicle. There is no doubt about it, especially with a base price of $ 35,700, for which you can get a mainstream, large-size SUV. Nevertheless, the small SUV from the Swedish manufacturer offers a whole range of high-end standard features such as several driving modes, a high-performance audio system, a 12.3-inch digital dashboard, a 9-inch touch screen and so on.
Therefore, it feels like the most equipped version of a mainstream SUV, whose average retail price is well over $40,000. Yet, as we are dealing with a luxury vehicle, there are always more as you add options and go for the higher trims.
The interior of the 2020 XC 40 has a contemporary Swedish styling and a very functional desing. The interior's light gives an impression of an upscale feeling even in the base trim. You get a pretty sizeable passenger space in both the front and rear seats. Of course, you will get more space with the XC60 and the XC90 trims. However, the XC 40 is very functional inside, and beautifully designed, especially when you look at the shift knob, crafted from Orrefors crystal.
The interior of the Volvo XC40 is very compact. For me it felt comfortable, but once I was transporting 3 adults in the back, and one of them said it could be a little bigger. But that wasn't everyone's case. Another person, sitting in the back, told me he was very comfortable. On the other hand, the SUV from the Swedish manufacturer offers us a fairly elegant dashboard and the quality of the materials that make up the interior is average.
The Exterior Design
When you hear the name Volvo, you can all imagine a car or SUV of good length and clean, elegant lines. For the 2020 XC40, it's quite the opposite and "it's not a bad thing since the latter does not give the impression of having passed through a compactor as is too often the case in this category," notes Jean-Sébastien Poudrier at Autobaine. The 2020 Volvo XC40's proportions are right and despite its aesthetic simplicity, it has something to catch the eye with a few key elements like its grille, headlights, taillights and design at the C-pillar. Volvo has also done an excellent job in terms of the choice of optional rims.
The XC40 is not Volvo's biggest trim, therefore the cargo space is comparable to that of some subcompact cars, but it's a sleek vehicle, packed with technology and a lot of fun to drive.
Armen Hareyan is the founder and the Editor in Chief of Torque News. He founded TorqueNews.com in 2010, which since then has been publishing expert news and analysis about the automotive industry. He can be reached at Torque News Twitter, Facebok, Linkedin and Youtube.
Volvo Owners to Inject $1.9 billion into Making Lotus Great Again
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How Does The Mustang Mach E Stack Up Against Other EVs, Like The Hot Hatch Polestar 2?
How The Volvo XC40 Recharge Compares To Other SUVs, Like The Jeep Wrangler 4xe
The Nice Folks At Consumer Reports Call The Polestar 2 EV Crossover A Failure
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Beeswax-jojoba oil mixture, 2 oz.
This spoon wax is what I use to treat all my work. It is tasteless, smells like honey, and won't change the coloring of the wood. Jojoba is a drought-tolerant plant native to the US southwest used in high grade cosmetics because it never goes rancid. It also makes a mean lip balm and hand salve. To order, please be in touch.