20 A qualitative densitometric HPTLC analysis was performed with methanolic extract for the development of characteristic

fingerprint profile, which may be used for quality evaluation and standardization of the drug. 10 μl of extract was spotted on pre-coated silica gel G60 F254 HPTLC plates (Merck) with the help of CAMAG Linomat V applicator. The plate was developed in glass twin trough chamber (20 cm × 10 cm) pre-saturated with mobile phase (Toluene: Ethyl acetate: Methanol: Glacial Acetic acid in the ratio 7.5:1.5:0.8:0.2). The plate was derivatized using methanolic H2SO4 and scanned using TLC Scanner 3 (CAMAG). The fruit is an indehiscent berry. It is an ellipsoid, obovoid or nearly cylindrical, slightly 5-sided, 7–10 cm long and 4–5 cm in diameter; capped by a thin, star-shaped calyx at the stem-end and tipped with five hair like floral remnants at the apex. Duvelisib price Crispy when unripe, the fruit turns from bright green to yellowish-green, ivory or nearly white when ripe and falls to the ground. The outer skin is glossy, very thin, soft and tender, and the pulp green, jelly-like and juicy (pH–2.4). There may be a few (6–7) flattened, disc-like seeds, 6 mm wide, smooth

and brown (Fig. 1B and C; Table 1). The T.S. of the fruit showed two distinct regions, exocarp and endocarp. Exocarp is the outermost layer of fruit made up of thin rectangular cells showing presence of simple and glandular trichomes and three to four layers of subepidermal collenchyma. In ripe fruits large lysigenously formed cavities are present in parenchyma with scattered conjoint, collateral and endarch AZD6738 vascular bundles. Endocarp cannot be differentiated in mature fruit as it disintegrates during ripening

of fruits (Fig. 1D–F). Powder microscopy shows the presence of simple and glandular trichomes, spiral thickening of vessels, tannin filled cells and fibres. (Fig. 1G–J). Ash of any organic material is composed of their non-volatile inorganic components. Controlled incineration of crude drugs results in an ash residue consisting of an inorganic material (metallic salts and silica). This value varies within fairly wide limits and is therefore very an important parameter for the purpose of evaluation of crude drugs.21 Therefore, percentage of the total ash, acid insoluble ash and water soluble ash were determined. The extraction of any crude drug with a particular solvent yields an extract containing different phyto-constituents. Extractive value is also useful for evaluation of crude drug, which gives an idea about the nature of the chemical constituents present in a crude drug and is useful for the estimation of specific constituents, soluble in that particular solvent used for extraction.16 Loss on drying is the loss of mass expressed as percent w/w.21 Results are tabulated in Table 2. The fluorescence character of powdered drug plays a vital role in the determination of quality and purity of the drug material.

However this observation in the murine model contrasts with documented evidence from the guinea pig Chlamydia caviae model of a primary genital tract infection in which chronic oviduct pathology was reported in only 12% of the animals, even though almost 80% were infected [66]. In humans, long-term chronic infections can develop after the primary infection [67] and the risk of pathology is known www.selleckchem.com/TGF-beta.html to

increase after repeated infections [5]. Thus the guinea pig model, with observed pathology following primary chlamydial infections and anatomy, and physiology similar to the human female genital tract, more closely resembles human chlamydial disease than the murine model. Choosing the most informative animal model to investigate CD4+ effector cell subsets elicited to combat C. trachomatis genital tract infections in humans will require prudence. Rather than AZD6738 mouse using the mouse strain, C. muridarum, several groups have used human C. trachomatis and shown that intravaginal inoculation of mice with this strain results a mild, self-limiting, lower reproductive tract infection with minimal ascension to the upper genital tract [68]. The eradication of C. trachomatis in the mouse is reportedly independent of indoleamine 2, 3-dioxygenase (IDO) [69] and yet this is a principle mechanism of protection against C. trachomatis infection in human cells, where IDO-catalyzed tryptophan degradation starves the chlamydial

inclusion of this amino acid [70]. Nevertheless, using this murine model, it has been established that to resolve genital chlamydial infection an influx of IFN-g-producing CD4+ Th1 cells is required [71] and [72] along with numerous host factors including matrix metalloproteinases (MMPs) such as MMP9 [73]. The host response to chlamydial infection is also proposed to directly damage mucosal tissues of the female genital tract. One hypothesis states that infected epithelial cells

secreting pro-inflammatory cytokines/chemokines to initiate pathogenesis (the cellular paradigm) whilst the second (immunological paradigm), as mentioned earlier in this paper, proposes that T-cell responses that are essential to resolve infection can also cause Bay 11-7085 tissue damage [46], [53] and [74]. The immunological paradigm for pathogenesis is supported by observations from both the guinea pig [75] and the non-human primate (NHP) [76] models of genital infection in which repeated oviduct infections cause rapid infiltration of CD4 and CD8 T cells to the infection site. Despite the fact that the majority of vaccine studies have been undertaken using the mouse model, there has also been a long history of non-human primate (NHP) models used in the study of chlamydial disease (dating back to 1936). The value of using NHPs as a model lies in their evolutionary closeness to Homo sapiens. NHPs have been particularly effective in the study of tubal pathology (pelvic inflammatory disease) (reviewed in [77]).

Amongst transporters present in the lungs (Bleasby et al., 2006), P-glycoprotein see more (P-gp, MDR1) and the organic cation/carnitine transporters (OCT and OCTN) have been detected in the human bronchial epithelium (Bosquillon, 2010). Although

the influence of lung transporters on drug pharmacokinetic profiles remain largely unknown, OCT/OCTN-mediated transport of inhaled therapeutic compounds in bronchial epithelial cell culture models has been suggested (Ehrhardt et al., 2005, Nakamura et al., 2010 and Mukherjee et al., 2012). On the other hand, there is considerable debate regarding the impact of P-gp on drug disposition in the lungs. Functional studies in rat models have demonstrated negligible transporter-mediated absorption of P-gp substrates either ex vivo ( Tronde et al., 2003 and Madlova et al., 2009) or in vivo ( Manford et al., 2005). In contrast, Francombe and colleagues have reported an increase in Rhodamine123 (Rh123) absorption from rat IPL in the presence of the P-gp potent inhibitor GF120918 in both the instillate and perfusate solutions ( Francombe et al., 2008). Similarly, studies that have investigated the functionality of P-gp in human bronchial epithelial cell layers are conflicting ( Bosquillon, 2010). Due to possible variations in substrate affinity for the human or

rat transporters, a reliable assessment of P-gp involvement in pulmonary drug absorption might only be achieved through a combination of in/ex vivo data in rats and in vitro permeability ABT 737 measurements in all both human and rat airway epithelial cell layers. An in vitro model of the rat respiratory epithelium would assist in the evaluation of the role of transporters as well as interspecies discrepancies in inhaled drug permeability. Importantly, bias in in vitro/in vivo absorption correlations resulting from transporter heterology, variable substrate

specificity and different pulmonary expression patterns in humans and rats would be minimised. This could improve the reliability of in vitro prediction and thus, guide the selection of drug candidates that progress to the late stages of pre-clinical development. Although a rat airway cell culture model is unlikely to replace drug testing in animals in the short term, it may nevertheless help reduce and refine the experimentation required. RL-65 is a rat airway (bronchial/bronchiolar) epithelial cell line that was isolated from 5 day old Sprague–Dawley rats (Roberts et al., 1990). This has been exploited to investigate cell-signalling pathways (Van Putten et al., 2001, Blaine et al., 2001, Wick et al., 2005, Bren-Mattison et al., 2005 and Nemenoff et al., 2008) or the epithelial–mesenchymal transition (Wang et al., 2009 and Felton et al., 2011) in airway epithelial cells preferentially to other cell lines due its non-cancerous origin and spontaneous immortalisation.

34 Grapes (Vitis vinifera) and wine are the most important sources of piceatannol. 35 It is also known as phytoalexins as it is produced in plants in stressed condition or against fungal attack. 34 It is a metabolite of resveratrol. It possesses an extra OH (hydroxyl) group at 3′ position in its structure. 36 It exhibits some properties that are analogous to resveratrol. It possesses more potent activity than resveratrol like good bioavailability,

low metabolization rate and high anti-oxidant activity. For showing its biological activity, it is required in a very small amount as compared SB431542 to resveratrol. Although there is a huge similarity between the biological activities of both the natural polyphenols, there are other properties of piceatannol like fetal hemoglobin induction which are still to be determined experimentally. It may be used as a new hope for the treatment of beta-thalassemic patients. Further studies should be done using this natural compound for checking its efficacy in HbF induction thereby making it clinically applicable for the treatment of beta-thalassemia. 35 Beta-thalassemic patients require regular blood transfusion for survival. They are unable to remove the free iron released from the transfused red blood Quizartinib cells. This excess iron gets deposited in the spleen, liver and endocrine organs. Iron accumulation leads to complications like diabetes, heart failure and finally

early death. Iron chelators form complex with tissue iron which is then excreted Parvulin in feces or

urine. Chelation therapy lessens iron-related complexities and improves quality of life. Some medicinal plants possessing iron chelating properties can also be used for the treatment of beta-thalassemia (Fig. 3).37 Deferoxamine (siderophore produced from Streptomyces griseus) is one of the most extensively used iron chelators used for treating transfusional iron overload in beta-thalassemic patients. It has been observed in thalassemic patients that deferoxamine possess a significant effect on long-term survival of the patients. Deferoxamine is the only chelator known which is responsible for the reversal of iron-induced heart failure. 38 and 39 Tetracarpidium conophorum (African walnut) extract possesses high chelating ability due to which it is used in industries as an iron chelating agent. It is used in the treatment of iron-overload disorders such as beta-thalassemia. Iron chelators from this plant extract lower iron availability in the blood circulation of thalassemic patients. 40 Wheatgrass (Triticum aestivum) belonging to Gramineae family, has been used since ancient times as a therapeutic for various diseases. 41 The crude extract of wheatgrass has been reported to contain iron chelating property. The oral intake of its juice may be helpful for beta-thalassemia. 42T. aestivum possess several beneficial effects in iron overload induced thalassemia like reduction in serum ferritin level and serum iron level in disease group.

Its contents are solely the responsibility of the authors and do not necessarily represent the official views of NIOSH-CDC. We would like to thank Mark Farfel, ScD, Carolyn Greene, MD, James L. Hadler, MD, MPH, Carey Maslow, PhD, Amanda Moy, MPH, Howard Alper, PhD, MS, Alice Welch, DrPH, RPh, and Margaret Millstone from the NYC Department of Health and Mental Hygiene, for their thoughtful comments, guidance, and support on this SCR7 datasheet manuscript. “
“Physical activity is an important, modifiable behavior for the prevention of non-communicable chronic diseases

(WHO). Epidemiological studies have shown that physical activity is associated with reduced risks of obesity, diabetes, cardiovascular disease, and other chronic diseases (Bize

et al., 2007 and Warburton et al., 2006). A growing number of studies have focused on the ecological context of physical activity (Sallis et al., 2008), i.e. the influence of the residential built environment on it (Trost et al., 2002). The built environment refers to the physical form of communities (Brownson et al., 2009), which has been operationalized according to 6 dimensions: residential density, street connectivity, accessibility to services and destinations, walking and cycling facilities, esthetic quality, and safety. There has been increasing evidence that the neighborhood built environment may influence residents’ physical Ulixertinib order activity, especially on transport-related physical activity (TRPA) and leisure-time physical activity (LTPA) (Fraser and Lock, 2011 and Owen et al., 2004). Chinese tuclazepam society has undergone rapid urbanization and urban sprawl, which have contributed to the decline of physical activity (Ng et al., 2009) and changes in residents’ physical activity pattern. For example,

the escalation of vehicle numbers (National Bureau of Statistics of China) is causing a reduction in traditional modes of TRPA (through walking, cycling and public transportation) in urban areas. Thus, it is critical to understand what and how built environment correlates with physical activity. Studies have been conducted in the U.S. (King et al., 2006), Australia (Humpel et al., 2002), Japan (Kondo et al., 2009), and Brazil (Hallal et al., 2010) to explore this possible relationship, yet few were carried out in China (Zhou et al., 2013). Furthermore, the demographic profile and SES (social-economic status) of the Chinese population could modify this relationships observed in other countries.

Consistent with published data [10], [11], [17] and [34], CaP acted as an adjuvant in this study and significantly enhanced CaP PCMC-induced antigen-specific IgG titres compared to soluble PCMCs. The adjuvant

effect of CaP and aluminium-based adjuvants has been attributed to their antigen depot effect [2] and [15]. However, the rate of antigen release from CaP PCMCs had no significant effect on the magnitude or duration of the antibody response and corroborates a growing body of evidence that the activity of traditional adjuvants is independent of a depot effect [35], [36] and [37]. It should be noted that no significant decrease in antigen-specific IgG titre was observed for Proteases inhibitor any formulation tested up to 84 d post-immunisation. However investigation of the antibody response for longer time periods might highlight differences between the different formulations. CaP PCMC promoted a decrease in antigen-specific IgG1:IgG2a ratio compared to Al(OH)3, indicating a more mixed Th1/Th2 immune response. Similar results have been obtained in other studies as a result of both CaP inclusion [17] and [38] and formulation into microparticle vaccines [39], [40] and [41]. As the adjuvant effect arising from surface modification of PCMC with CaP was independent of CaP loading, we hypothesised that the morphology

of CaP PCMCs may be important for their Lumacaftor adjuvant activity. PCMCs are of suitable size and morphology to be phagocytosed by immune

cells [42] and phagocytosis of latex microspheres by monocytes else promotes their differentiation to functional dendritic cells and subsequent immune priming in the draining lymph node [43]. Formulation into PCMCs without CaP enhanced phagocytosis of BSA-FITC by J774.2 cells, possibly due to enhanced cell function arising from the l-glutamine released from the core component of the soluble PCMCs [30], [31], [32] and [33]. However, the phagocytosis of BSA-FITC was clearly further enhanced by formulation into CaP PCMCs. Thus, CaP PCMCs may exert their adjuvant effect, at least in part, through enhanced uptake of antigen by tissue phagocytes and subsequent enhancement of immune priming. However, further studies are needed to determine the precise mechanism by which CaP PCMCs exert their adjuvant effect in vivo. Combined with published data [5] and [7], our results indicate that CaP PCMCs represent a useful platform by which to progress future vaccine formulation. SJ performed PCMC preparation, SEM analysis and determination of antigen-specific IgG, IgG1 and IgG2a titres pertaining to PCMCs loaded with DT, CyaA* and BSA. CA performed all in vivo experiments. DK prepared PCMCs loaded with BSA-FITC, analysed PCMC uptake by flow cytometry and stained cells for CLSM.

1 × 106 K562 cells were incubated for 24 h and then irradiated (1 J/cm2) in HBSS with or without the test compounds. After 24 h from irradiation, cells were fixed with ice-cooled ethanol (70% v/v), treated overnight with RNAse A (0.1 mg/mL) in phosphate saline buffer and finally stained with propidium iodide (PI, 0.1 mg/mL). Samples were analyzed on a BD FACS Calibur flow cytometer collecting 10,000 events. Results of cell-cycle analysis were examined

using WinMDI 2.9 (Windows Multiple Document Interface for Flow Cytometry) [20]. K562 cells (300,000 cells/mL) were seeded in 24-well microplate CB-839 datasheet and incubated for 24 h prior irradiation. After medium removal, 1 mL of the drug solution was added to each well, incubated at 37 °C for 30 min

and then irradiated (1 J/cm2). After irradiation, the solution was replaced with complete medium and the plates were incubated for 24 h. Cells were collected by centrifugation and re-suspended in 1 μM JC-1 (5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazol-carbocyanine) solution PFT�� in HBSS or in 100 nM NAO (10-N-nonyl acridine orange) solution in RPMI medium. The cytofluorimetric analysis (BD FACS Calibur flow cytometer) was performed collecting green (FL1) and orange (FL2) fluorescence for JC-1 staining and only the green one (FL1) for NAO staining in at least 10,000 events for each sample [22] and [23]. Solutions of derivatives in methanol were irradiated in a quartz cuvette with different UV-A doses (0, 8, 16 and 32 J/cm2). After the irradiation, the solution was lyophilized, suspended in a known volume of methanol and stored at −20 °C. Concentrations of unknown photoproduct mixtures in this paper were expressed as if the initial psoralen was not photodegraded. RNA was isolated from K562 cells and measured by reverse transcription quantitative

real-time polymerase chain reaction (RT-qPCR) as described [24] using gene-specific double fluorescence labeled probes in an ABI Prism 7700 Sequence Detection System version 1.7.3 (Applied Biosystems). The following primer and probe sequences were used: α-globin forward primer, 5′-CAC GCG CAC AAG CTT CG-3′; α-globin reverse primer, 5′-AGG GTC ACC AGC AGG CAG T-3′; α-globin probe, 5′-FAM-TGG ACC CGG TCA ACT TCA AGC TCC T-TAMRA-3′; γ-globin forward until primer, 5′-TGG CAA GAA GGT GCT GAC TTC-3′; γ-globin reverse primer, 5′-TCA CTC AGC TGG GCA AAG G-3′; γ-globin probe, 5′-FAM-TGG GAG ATG CCA TAA AGC ACC TGG-TAMRA-3′. The kit for quantitative RT-PCR for ζ-globin mRNA and ε-globin mRNA were from Applied Biosystems (ζ-globin mRNA: Hs00923579_m1; ε-globin mRNA: Hs00362216_m1). The fluorescent reporter and the quencher were 6-carboxyfluorescein (FAM) and 6-carboxy-N,N,N′,N′-tetramethylrhodamine (TAMRA), respectively. For real-time PCR, the reference gene was 18S; this probe was fluorescent-labeled with VIC (Applied Biosystems) [24] and [25]. Unless indicated otherwise, results are presented as mean ± SEM.

The nucleotide sequences of the HA and selleck products NA of SH1 and AH1 were downloaded from the GISAID Epiflu database (accession numbers EPI439486 and EPI439507, respectively). Gene synthesis was conducted by GeneArt

(Life Technologies, Carlsbad, CA). SH1 and AH1 HA and NA sequences were subcloned into the ambisense rescue plasmid pDZ for rescue of recombinant influenza viruses. Additional recombinant PR8 virus (7:1) were generated that expressed the HA of the H7 Eurasian lineage virus A/mallard/NL/12/00 (H7N3; PR8:malNL00), or the HA of A/chicken/Jalisco/12283/12 (H7N3; PR8:chickJal12) which was genetically modified to remove the multibasic cleavage site. An additional recombinant PR8 viruses was included that expressed a chimeric cH7/3 HA in which the globular head domain was derived from the H7 North American lineage virus A/mallard/Alberta/24/01 (H7N3; PR8:malAlb01) on an H3 stalk [21] and [22]. Viruses were propagated in 8- to 10-day-old specific pathogen-free embryonated chicken eggs (Charles River Laboratories) for 48 h at 37 °C and virus was titred on MDCK cells in the presence of tosyl phenylalanyl chloromethyl ketone (TPCK) treated trypsin. Synthesised SH1 and AH1 HA genes (GISAID Epiflu database accession numbers EPI439486 and EPI439507, respectively) and the matrix protein (M1) gene from strain A/Udorn/307/72 (H3N2) (GenBank: DQ508932.1),

synthesised by Sloning (Puchheim,

Germany), were cloned as previously described [17]. VLPs consisting of the respective learn more H7 HA (either AH1 or SH1) and the matrix protein (M1) from the unrelated H3N2 subtype were produced by baculovirus infection of insect cells as described before [17]. Empty VLPs consisting of M1 only were prepared to be used as a negative control. Briefly, the synthetic genes were cloned into a modified pFastBacDual baculovirus transfer vector and recombinant bacmids were constructed using the Bac-to-Bac System (Invitrogen, Carlsbad, CA). Recombinant baculovirus over was rescued from Sf9 cells and amplified. VLPs were expressed in High Five cells using Fernbach flasks incubated at 27 °C. Cells were infected with the recombinant baculoviruses at a multiplicity of infection of approximately 5 and culture supernatant was harvested 4 days post infection by low-speed centrifugation (3.000 rpm, 10 min). VLPs were partially purified and concentrated using a 30% (w/v) sucrose cushion in phosphate buffered saline (PBS) and the pellet was resuspended in PBS and stored at 4 °C. To quantify the HA content of the VLPs, different concentrations of VLP samples were compared to known concentrations of recombinant His-tag purified SH1-HA containing a T4 foldon trimerisation domain [23]. VLP and His-tag HA were separated by SDS-PAGE using 4–12% gradient polyacrylamide gels (Invitrogen, Carlsbad, CA).

Activity interference was also recorded in the diaries daily using Item 5 from the 12-Item Short-Form Health Survey (Ware et al 1996), a 5-point scale anchored by ‘not at all’ through to ‘extreme interference’. To ensure completeness of follow-up, data from the diaries were collected by telephone interview at weekly intervals for the first four weeks, then monthly or until recovery for the subsequent eight Selleck Wnt inhibitor weeks (84 days in total). At

three months, a telephone exit interview was conducted at which the Neck Disability Index (Vernon and Mior 1991) was administered and pain scores were collected. Primary outcome: The primary outcome was the time taken from commencement of treatment to recovery from the episode of neck pain. The day of recovery from the episode of neck pain was defined as the first day of seven consecutive days on which the patient rated the intensity of their average daily neck pain as < 1 on the numerical rating scale from 0 to 10. Secondary outcomes: Secondary outcomes included time to recovery of normal activity as well as pain (numerical rating scale 0–10) and disability GSK1349572 concentration (Neck Disability Index scale 0–50) scores at

three months. Time to recovery of normal activity was defined as the first day of seven consecutive days in which the participant rated the degree of interference ‘not at all’. We examined 22 putative prognostic factors. Eight demographic variables were examined: age, gender, level of education, employment status, change of employment status due to neck pain, smoking habit, whether a compensation claim for neck pain had been lodged, and self-rated general health. Level of education was determined using items from the Australian Census 2001 (Trewin 2000). Employment status was determined using categories described by

Kenny et al (2000). Self-rated general health was measured using Item 1 of the 12-Item Short-Form Health Survey (SF-12). The 14 clinical variables examined were: pain intensity on the 0–10 numerical rating scale, duration of neck pain, disability measured by the Neck Disability Index from 0 (none) to 50 (worst), the physical (PCS) and mental health (MCS) component summary scales of the SF-12, presence of concomitant symptoms (upper limb pain, headache, upper back pain, lower back pain, dizziness and nausea), past history of neck pain, previous sick leave for isothipendyl neck pain, and use of analgesics. The clinical course of the episode of neck pain was described using Kaplan-Meier survival curves and using descriptive statistics. Prognostic factors were evaluated using separate prognostic models for recovery from the episode of neck pain and disability at 3 months. The first stage involved examination of the univariate relationship between the outcome and each prognostic variable, using Cox regression (for time to recovery), and linear regression (for disability at 3 months). Variables with significant associations (p < 0.

Once approved, the new recommendations are distributed in an official letter or in a revised edition of the immunization reference manual to all public health facilities

in the country and posted online on the website of the DDC. The new recommendations are also announced in annual refresher ON-01910 concentration courses conducted by the national EPI for all health workers involved in immunizations. For many years, the ACIP has played a key role in guiding decisions related to vaccine use and immunization in Thailand and the Committee is considered an important factor in the success of the country’s national immunization program. There are a number of factors contributing to the success of the Committee. These include: its formal establishment by the Minister of Public Health; the multi-disciplinary expertise among its members; and the fact that the Secretariat consists of those responsible for implementing the national immunization program. However, the ACIP has a number of limitations which could be addressed to further strengthen the Committee and how it functions. These limitations and possible areas of improvement include the following: (1) There are no regulations or laws stipulating that all immunization-related policy decision must first be considered by the ACIP. There have therefore been instances in which

new immunization policies were

enacted without consideration by the Committee. The authors state selleck products that they have no conflict of interest. We wish to acknowledge Dr. Sujarti Jetanasen, Dr. Prayura Kunasol, Dr. Supamit Chunsuttiwat, and Denise DeRoeck. The three authors of this paper are all members of the Thai ACIP. “
“Figure options Download full-size image Download as PowerPoint slide This supplement is dedicated to the late Professor V. Borovick. Professor Borovick died at the age of 67 on August 25, 2009, in Serpukhov, Russia, before he could see this publication come to fruition. A great loss comes with Professor Borovick’s passing. It is with a renewed sense of purpose that we dedicate this supplement of the journal to him and his lifelong efforts to use science and technology as a uniting very force in international relations. Professor Borovick was an outstanding scientist in the field of infectious diseases, pathogenesis, immuno- and biochemistry, medical biotechnology, veterinary medicine, and agriculture. Those who knew Professor Borovick remember, with tremendous admiration, his commandeering one of the most exciting and successful post-Cold War international collaborations of scientific activity between Russian ministries and government agencies, private organizations, academic institutions, and the U.S. government agencies. His partners included U.S.