Barriers to opioid use disorder treatment among people who use drugs in the rural United States: A qualitative, multi-site study.

BACKGROUND: In 2020, 2.8 million people required substance use disorder (SUD) treatment in nonmetropolitan or 'rural' areas in the U.S. Among this population, only 10% received SUD treatment from a specialty facility, and 1 in 500 received medication for opioid use disorder (MOUD). We explored the context surrounding barriers to SUD treatment in the rural United States. METHODS: We conducted semi-structured, in-depth interviews from 2018 to 2019 to assess barriers to SUD treatment among people who use drugs (PWUD) across seven rural U.S. study sites. Using the social-ecological model (SEM), we examined individual, interpersonal, organizational, community, and policy factors contributing to perceived barriers to SUD treatment. We employed deductive and inductive coding and analytical approaches to identify themes. We also calculated descriptive statistics for participant characteristics and salient themes. RESULTS: Among 304 participants (55% male, mean age 36 years), we identified barriers to SUD treatment in rural areas across SEM levels. At the individual/interpersonal level, relevant themes included: fear of withdrawal, the need to "get things in order" before entering treatment, close-knit communities and limited confidentiality, networks and settings that perpetuated drug use, and stigma. Organizational-level barriers included: strict facility rules, treatment programs managed like corrections facilities, lack of gender-specific treatment programs, and concerns about jeopardizing employment. Community-level barriers included: limited availability of treatment in local rural communities, long distances and limited transportation, waitlists, and a lack of information about treatment options. Policy-level themes included insurance challenges and system-imposed barriers such as arrest and incarceration. CONCLUSION: Our findings highlight multi-level barriers to SUD treatment in rural U.S. communities. Salient barriers included the need to travel long distances to treatment, challenges to confidentiality due to small, close-knit communities where people are highly familiar with one another, and high-threshold treatment program practices. Our findings point to the need to facilitate the elimination of treatment barriers at each level of the SEM in rural America.

Increased innervation of the vulval vestibule in patients with vulvodynia.

BACKGROUND: Vulval vestibulitis is a condition characterized by the sudden onset of a painful burning sensation, hyperalgesia, mechanical allodynia, and occasionally pruritus, localized to the region of the vulval vestibulus. It is considered the commonest subset of vulvodynia. Pain precipitated in the absence of nociceptor stimuli might be triggered by previous peripheral nerve injury, or by the release of neuronal mediators, which set off inappropriate impulses in nonmyelinated pain fibres sensitizing the dorsal horn neurones. The pathophysiology of vulval vestibulitis is still unclear. OBJECTIVES: The objective of this study was to evaluate the nerve fibre density and pattern, in specimens of vulval vestibulus, in normal subjects and in patients with vestibulitis, and provide objective diagnostic criteria for this condition. Methods Twelve patients with a history of the vestibulitis type of vulvodynia, and eight normal subjects underwent biopsy of the posterior wall of the vulval vestibule. Quantitative immunohistochemistry was performed, using antisera to the general neuronal marker protein gene product (PGP) 9.5, and to the neuropeptide calcitonin gene-related peptide (CGRP), on 15- microm sections. RESULTS: There was a statistically significant increase of density and number of PGP 9.5 immunoreactive in the papillary dermis of patients with vulvodynia of the vestibulitis type, compared with those of controls. However, the distribution pattern of the innervation showed no significant change. There were no significant differences in CGRP staining between patients and controls. CONCLUSIONS: It is concluded that the increase of PGP 9.5 immunoreactive nerve fibres, in patients with vulvodynia, may be either secondary to nerve sprouting, or may represent neural hyperplasia. Increased innervation may be applied as an objective diagnostic finding in vulval vestibulitis syndrome.

Effect of transgenic plants expressing high levels of a tobacco anionic peroxidase on the toxicity of Anagrapha falcifera nucleopolyhedrovirus to Helicoverpa zea (Lepidoptera: Noctuidae).

Wild type and corresponding transgenic tomato (Lycopersicon esculentum Miller) and two tobacco (Nicotiana spp.) plants that express high levels of a tobacco anionic peroxidase were used to determine what type of interactions occurred between peroxidase altered plant chemistry and the baculovirus Anagrapha falcifera nucleopolyhedrovirus (AfMNPV) for control of neonate corn earworms, Helicoverpa zea (Boddie). Transgenic plants expressed approximately five to 400 times higher peroxidase activity than corresponding tissues of wild type plants. The H. zea larvae typically fed 1.5 times less on transgenic compared with wild type leaf disks. There was only one experiment (of three with tomato leaves) where the larvae that fed on transgenic leaves were less susceptible to the virus based on nonoverlapping 95% confidence intervals for LC50 values. When the exposure dose was corrected for reduced feeding on the transgenic leaf disks, the insecticidal activity of the virus was not significantly different for larvae fed on transgenic versus wild type plants. Eight other experiments (with tomato and two species of tobacco) indicated either no significant effect or enhanced susceptibility (when corrected for feeding rates) to the virus of larvae fed on the transgenic leaves. These results indicate enhanced insect resistance in plants expressing high levels of a specific anionic peroxidase may be compatible with applications of AfMNPV. Potential reasons for this compatibility are discussed.

Local and systemic expression of basic fibroblast growth factor in a patient with familial glomangioma.

Glomangiomas are rare cutaneous tumours composed of glomus cells, which are modified smooth muscle cells. The aetiology of this condition is thought to involve a mutation in a novel gene acting to regulate angiogenesis. We report a patient from a large family with three generations affected by familial multiple glomangiomas. We hypothesized that the growth factors basic fibroblast growth factor and vascular endothelial growth factor, which stimulate/regulate angiogenesis could be involved in the pathogenesis of these lesions. Therefore, using enzyme-linked immunosorbent assays and immunohistochemistry, respectively, we measured systemic and tissue levels of these growth factors in a patient with familial glomangiomas. In addition, we investigated endothelial mitogenicity of the patient's serum as a functional assay of systemic growth factor activity.

BFIT, a unique acyl-CoA thioesterase induced in thermogenic brown adipose tissue: cloning, organization of the human gene and assessment of a potential link to obesity.

We hypothesized that certain proteins encoded by temperature-responsive genes in brown adipose tissue (BAT) contribute to the remarkable metabolic shifts observed in this tissue, thus prompting a differential mRNA expression analysis to identify candidates involved in this process in mouse BAT. An mRNA species corresponding to a novel partial-length gene was found to be induced 2-3-fold above the control following cold exposure (4 degrees C), and repressed approximately 70% by warm acclimation (33 degrees C, 3 weeks) compared with controls (22 degrees C). The gene displayed robust BAT expression (i.e. approximately 7-100-fold higher than other tissues in controls). The full-length murine gene encodes a 594 amino acid ( approximately 67 kDa) open reading frame with significant homology to the human hypothetical acyl-CoA thioesterase KIAA0707. Based on cold-inducibility of the gene and the presence of two acyl-CoA thioesterase domains, we termed the protein brown-fat-inducible thioesterase (BFIT). Subsequent analyses and cloning efforts revealed the presence of a novel splice variant in humans (termed hBFIT2), encoding the orthologue to the murine BAT gene. BFIT was mapped to syntenic regions of chromosomes 1 (human) and 4 (mouse) associated with body fatness and diet-induced obesity, potentially linking a deficit of BFIT activity with exacerbation of these traits. Consistent with this notion, BFIT mRNA was significantly higher ( approximately 1.6-2-fold) in the BAT of obesity-resistant compared with obesity-prone mice fed a high-fat diet, and was 2.5-fold higher in controls compared with ob/ob mice. Its strong, cold-inducible BAT expression in mice suggests that BFIT supports the transition of this tissue towards increased metabolic activity, probably through alteration of intracellular fatty acyl-CoA concentration.

Biotic and abiotic factors limiting efficacy of Bt corn in indirectly reducing mycotoxin levels in commercial fields.

Incidence of insect damage, and association of insect damage with mycotoxigenic corn ear molds and mycotoxins was examined in commercial fields of Bt and non-Bt hybrids of different backgrounds in Illinois in 1998 and 1999. Nearly 50% Helicoverpa zea (Boddie) infestation sometimes occurred in Bt hybrids that express high levels of the protein in silks and kernels. Damage by European corn borer, Ostrinia nubilalis Hübner, was uncommon, even in non-Bt ears. Levels of total fumonisins were generally less (15- to 1.8-fold) in Bt versus non-Bt hybrids at the same site, with some significant differences. There were several instances where there were no significant differences in fumonisin levels between low/no Bt kernel hybrids and Bt hybrids that produced high levels of the protein in the kernel and silk tissue. However, significant correlations were often noted between numbers of insect-damaged kernels and total fumonisin levels, especially in 1998, suggesting in these cases that reducing insect damage was still reducing fumonisin levels. There was variability between the correlation coefficient for numbers of insect damaged kernels and fumonisin levels at different sites for the same year, different hybrids at the same site, and the same hybrid for different years. Although reductions in fumonisins in Bt hybrids were more limited than reported in the past, planting the Bt hybrids still appears to be a useful method for indirectly reducing mycotoxins in corn ears.

Maize rhm1 resistance to Bipolaris maydis is associated with few differences in pathogenesis-related proteins and global mRNA profiles.

The maize rhm1 mutant resists Bipolaris maydis, the causal agent of Southern corn leaf blight, by producing small necrotic lesions surrounded by chlorotic haloes. The rhm1 and wild-type lesions contain viable fungus in equal frequency, but fungal sporulation was markedly inhibited on rhm1. The levels of the pathogenesis-related (PR) proteins chitinase, PR1, and peroxidase differ little between rhm1 and wild type, with or without B. maydis inoculation. The global mRNA profiles surveyed revealed hundreds of cDNA fragments that were twofold or more induced or suppressed in rhm1 and wild-type plants following B. maydis inoculation. Nonetheless, between rhm1 and wild type, only 0.4 to 0.7% of the cDNA fragments were expressed differentially by twofold or more. Among the up-regulated genes in rhm1 was beta-glucosidase glu1, which prompted a test of whether rhm1 resistance depends upon the antimicrobial compound 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one or other hydroxamic acids whose glucosyl conjugates are preferred substrates for the Glu1 enzyme. Double mutants of rhm1 and bx1, a hydroxamic acid-deficient mutant, indicate that rhm1 resistance is hydroxamic acid independent. The rhm1 resistance presently appears to operate via a mechanism unlike those of previously described resistance genes.

Isolation and characterization of kinase interacting protein 1, a pollen protein that interacts with the kinase domain of PRK1, a receptor-like kinase of petunia.

Many receptor-like kinases have been identified in plants and have been shown by genetic or transgenic knockouts to play diverse physiological roles; however, to date, the cytosolic interacting proteins of relatively few of these kinases have been identified. We have previously identified a predominantly pollen-expressed receptor-like kinase of petunia (Petunia inflata), named PRK1, and we have shown by the antisense RNA approach that it is required for microspores to progress from the unicellular to bicellular stage. To investigate the PRK1-mediated signal transduction pathway, PRK1-K cDNA, encoding most of the cytoplasmic domain of PRK1, was used as bait in yeast (Saccharomyces cerevisiae) two-hybrid screens of pollen/pollen tube cDNA libraries of petunia. A protein named kinase interacting protein 1 (KIP1) was found to interact very strongly with PRK1-K. This interaction was greatly reduced when lysine-462 of PRK1-K, believed to be essential for kinase activity, was replaced with arginine (the resulting protein is named PRK1-K462R). The amino acid sequence of KIP1 deduced from full-length cDNA contains an EF-hand Ca(2+)-binding motif and nine predicted coiled-coil regions. The yeast two-hybrid assay and affinity chromatography showed that KIP1 interacts with itself to form a dimer or higher multimer. KIP1 is present in a single copy in the genome, and is expressed predominantly in pollen with a similar temporal pattern to PRK1. In situ hybridization showed that PRK1 and KIP1 transcripts were localized in the cytoplasm of pollen. PRK1-K phosphorylated KIP1-NT (amino acids 1--716), whereas PRK1-K462R only weakly phosphorylated KIP1-NT in vitro.

Vertilecanins: new phenopicolinic acid analogues from Verticillium lecanii.

Five new phenopicolinic acid analogues (1-5) have been isolated from solid-substrate fermentation cultures of Verticillium lecanii. The most abundant component (vertilecanin A; 1) displays antiinsectan activity against Helicoverpa zea. These compounds were obtained by chromatographic fractionation of the EtOAc culture extract and identified by analysis of NMR and MS data. The known fungal metabolites 2-decenedioic acid and 10-hydroxy-8-decenoic acid were also isolated from these cultures.

Changes in root cap pH are required for the gravity response of the Arabidopsis root.

Although the columella cells of the root cap have been identified as the site of gravity perception, the cellular events that mediate gravity signaling remain poorly understood. To determine if cytoplasmic and/or wall pH mediates the initial stages of root gravitropism, we combined a novel cell wall pH sensor (a cellulose binding domain peptide-Oregon green conjugate) and a cytoplasmic pH sensor (plants expressing pH-sensitive green fluorescent protein) to monitor pH dynamics throughout the graviresponding Arabidopsis root. The root cap apoplast acidified from pH 5.5 to 4.5 within 2 min of gravistimulation. Concomitantly, cytoplasmic pH increased in columella cells from 7.2 to 7.6 but was unchanged elsewhere in the root. These changes in cap pH preceded detectable tropic growth or growth-related pH changes in the elongation zone cell wall by 10 min. Altering the gravity-related columella cytoplasmic pH shift with caged protons delayed the gravitropic response. Together, these results suggest that alterations in root cap pH likely are involved in the initial events that mediate root gravity perception or signal transduction.

The effect of fungal ribosome inactivating proteins upon feeding choice in C. freemani, and indications of a mutualistic relationship with A. restrictus. Environmental mycology.

Carpophilus freemani beetles' feeding on the fungus Aspergillus nidulans was substantially inhibited when A. nidulans was transformed and induced to secrete the ribosome inactivating protein, restrictocin (genetic source: Aspergillus restrictus). No inhibition of feeding was observed when A. nidulans was transformed and induced to produce an inactive form of restrictocin with a single amino-acid substitution in the active site. Similarly, there was no inhibition of feeding upon transgenic strains when the production of restrictocin was not induced. Feeding inhibition of C. freemani by restrictocin requires that the ribonuclease be active and is not due to other characteristics of the protein or the transgenic host fungus.

Multiacreage evaluation of aerially applied adherent malathion granules for selective insect control and indirect reduction of mycotoxigenic fungi in specialty corn.

Aerially applied adherent corn flour granules containing 1% malathion were more often as, or more, effective than 15% chlorpyrifos (Lorsban 15G) granules in controlling caterpillars and sap beetles in high amylose corn in 1997 than 1996. Use of malathion granules corresponding closely in size to chlorpyrifos granules in the second year of the study apparently increased relative efficacy. Significantly less corn borer damage occurred on plants (1996) or ears (1997) within 2 wk of application for both types of insecticide granules compared with untreated plots. In 1997, there were sixfold fewer milk stage ears with more than 20 kernels damaged per ear in the malathion-treated plots compared with chlorpyrifos-treated plots, and severity of caterpillar damage was also less in malathion versus chlorpyrifos-treated plots at harvest. Control of beetles (corn rootworm adults and sap beetles) for both treatments was less effective compared with caterpillars. Significant corn rootworm adult control was noted for both chlorpyrifos and malathion in 1996 and significant sap beetle control was noted for the malathion granules in 1997. Significantly fewer live lady beetles, and more dead lady beetles were present in chlorpyrifos-treated plots compared with malathion-treated or untreated plots in 1996. The incidence and severity of Fusarium mold on ears at harvest was often indirectly reduced by both malathion treatments and chlorpyrifos treatments, with the malathion treatment significantly better than the chlorpyrifos treatment in one case.

New cyclic peptide and bisindolyl benzenoid metabolites from the sclerotia of Aspergillus sclerotiorum.

Scleramide (1), a new cyclic hexapeptide, and a new oxidized bisindolyl benzenoid derivative (2' '-oxoasterriquinol D methyl ether, 2) were isolated from extracts of the sclerotia of Aspergillus sclerotiorum (NRRL 5167). The structures of these compounds were determined by analysis of 1D and 2D NMR experiments.

Isoform patterns of chitinase and beta-1,3-glucanase in maturing corn kernels (Zea mays L.) associated with Aspergillus flavus milk stage infection.

Isoform patterns of chitinase and beta-1,3-glucanase of maturing kernels of yellow dent corn (Pioneer 3394) infected with Aspergillus flavus at the milk stage were investigated through polyacrylamide gel electrophoresis (PAGE). Proteins on the sodium dodecyl sulfate (SDS) gel with an apparent molecular mass range of 23-46 kDa were differentially present in the kernels infected with both aflatoxin-producing and non-aflatoxin-producing strains of A. flavus. From in-gel (native PAGE) enzyme activity assays, three bands corresponding to chitinase isoforms and two bands corresponding to beta-1,3-glucanase isoforms were detected in the infected kernels. One chitinase isoform of 29 kDa was present only in the infected kernels, and another one of 28 kDa was present in both infected and noninfected kernels. They were judged to be acidic on the basis of their migration on an acrylamide isoelectric focusing (IEF) gel. For the beta-1,3-glucanase, one isoform of 35 kDa was present in both infected and noninfected kernels, but another one, a 33 kDa isoform, was present only in the infected kernels. Both acidic and basic beta-1,3-glucanase isoforms were detected in the IEF gel. The results of this study are the first to demonstrate patterns of enhanced or inducible proteins in maturing corn kernels in response to A. flavus infection at the milk stage. The results also indicate that only particular isoforms of the two hydrolytic enzymes are involved in the maturing corn kernels infected at the milk stage with A. flavus.

Cloning and characterization of IL-17B and IL-17C, two new members of the IL-17 cytokine family.

IL-17 is a T cell-derived cytokine that may play an important role in the initiation or maintenance of the proinflammatory response. Whereas expression of IL-17 is restricted to activated T cells, the IL-17 receptor is found to be widely expressed, a finding consistent with the pleiotropic activities of IL-17. We have cloned and expressed two novel human cytokines, IL-17B and IL-17C, that are related to IL-17 ( approximately 27% amino acid identity). IL-17B mRNA is expressed in adult pancreas, small intestine, and stomach, whereas IL-17C mRNA is not detected by RNA blot hybridization of several adult tissues. No expression of IL-17B or IL-17C mRNA is found in activated T cells. In a survey of cytokine induction, IL-17B and IL-17C stimulate the release of tumor necrosis factor alpha and IL-1beta from the monocytic cell line, THP-1, whereas IL-17 has only a weak effect in this system. No induction of IL-1alpha, IL-6, IFN-gamma, or granulocyte colony-stimulating factor is found in THP-1 cells. Fluorescence-activated cell sorter analysis shows that IL-17B and IL-17C bind to THP-1 cells. Conversely, IL-17B and IL-17C are not active in an IL-17 assay or the stimulation of IL-6 release from human fibroblasts and do not bind to the human IL-17 receptor extracellular domain. These data show that there is a family of IL-17-related cytokines differing in patterns of expression and proinflammatory responses that may be transduced through a cognate set of cell surface receptors.

Indirect reduction of ear molds and associated mycotoxins in Bacillus thuringiensis corn under controlled and open field conditions: utility and limitations.

In 1995, ears of a experimental inbred (CG59-2) containing a synthetic Bacillus thuringiensis Cry IA (b) gene driven by PEPC, pith and pollen promoters and artificially infested with Ostrinia nubilalis (Hübner) larvae in small plot studies were free from insect damage, whereas 40-50% of the corresponding non-Bt inbred ears were damaged. Bt inbred ears that were inoculated with Aspergillus flavus Link and Fusarium proliferatum T. Matsushima (Nirenberg) or exposed to natural mold inoculum after infestation with O. nubilalis were free of visible signs of mold, as compared with approximately 30-40% of the non-Bt ears similarly treated. Results in 1996 using the same inbred with a single allele dose of the Bt gene showed similar trends. Mean total fumonisin levels for non-Bt versus Bt inbred ears were not significantly different (2.8 versus 0.8 ppm, respectively) in 1996. In paired hybrid studies run in 0.4-ha (1-acre) fields, an event 176 Bt hybrid had significantly lower amounts of damage and signs of Fusarium spp. mold, but not fumonisin, compared with a corresponding non-Bt hybrid from 1996 to 1998. However, two hybrid pairs that contained either MON810 or Bt11 constructs examined in similar fields at the same site had lower levels of fumonisin in both 1997 (30- to 40-fold) and 1998. High intrafield variability in insect infestation and presence of Helicoverpa zea (Boddie) in Bt hybrids was apparently responsible for fewer significant differences in fumonisin levels in 1998. Similar trends for all three hybrid pairs were noted in small plot trials at another site. Incidence of other ear pests or insect predators varied as much among non-Bt hybrids as they did for Bt/non-Bt hybrid pairs.

Dusky sap beetles (Coleoptera: Nitidulidae) and other kernel damaging insects in Bt and non-Bt sweet corn in Illinois.

Bt and non-Bt sweet corn hybrids (Rogers 'Empire' Bt and non-Bt, respectively) were compared for distribution of kernel damaging insect pests in central Illinois in 1998 and 1999. The occurrence and damage by caterpillars [primarily Helicoverpa zea (Boddie)] were reduced by at least 80% in each year for the Bt compared with the non-Bt hybrid. However, the incidence of sap beetle adults (primarily Carpophilus lugubris Murray) was higher, and larvae, lower for the Bt versus non-Bt in 1999. The incidence of ears with more than five kernels damaged by sap beetles was higher for the Bt compared with non-Bt hybrid in 1998 (13.8 versus 5.5%), but nearly equivalent in 1999 (15.3 versus 15.1%, respectively). Distribution of predators on plants (primarily Coccinelidae) and harvested ears (primarily Orius spp.) were not significantly different on Bt versus non-Bt hybrids. Ears with husks flush with the ear tip or with ear tips exposed had significantly higher sap beetle damage for both hybrids, and the Bt hybrids had significantly higher incidence of exposed ear tips in both years. Sap beetle numbers determined by scouting were often proportional to numbers of beetles captured in baited traps, increasing and decreasing at about the same time. However, values determined with traps were typically less variable than when scouted, and time of sampling was typically four times more rapid for each trap than for each 10 plant scout sample when measured in 1999.