GeoHealth Perspectives on Integrated, Coordinated, Open, Networked (ICON) Science.

GeoHealth research both characterizes and predicts problems at the nexus of earth and human systems like climate change, pollution, and natural hazards. While GeoHealth excels in the area of integrated science, there is a need to improve coordinated and networked efforts to produce open science to enable environmental justice. There is a need to resource and empower frontline populations that are disproportionately marginalized by environmental injustice (i.e., the unequal protection from environmental harms and lack of access and meaningful engagement in decision making for a healthy environment; EPA, 2022, https://www.epa.gov/environmentaljustice). GeoHealth practice has the opportunity to advance environmental justice or the "fair treatment and meaningful involvement of all people regardless of race, color, national origin, or income" with respect to how research and collaboration of GeoHealth professionals supports the "development, implementation, and enforcement of environmental laws, regulations, and policies" that produce equal protection from environmental and health hazards and access to the decision making for a health environment (EPA, 2022, https://www.epa.gov/environmentaljustice). Here we highlight barriers and opportunities to apply an equity-centered ICON framework to the field of GeoHealth to advance environmental justice and health equity.

Resolution of persistent oral papillomatosis in a dog after treatment with a recombinant canine oral papillomavirus vaccine.

This report describes a 16-month-old female, otherwise seemingly healthy, Siberian husky dog with severe oral papillomatosis that did not regress spontaneously and was refractory to surgical treatment over a 6-month period. Regression of the papillomas was achieved by administering a series of experimental vaccinations starting at the time of the last surgery. The vaccine consisted of systemically administered canine oral papillomavirus major coat protein L1 that has been shown to self-assemble into virus-like particles. They cause a humoral response that has been shown to prevent the onset and development of papillomas. In this case, however, following unsuccessful surgical treatment, the vaccine acted therapeutically, causing the papillomas that had regrown to shrink. No side-effects were noted.

Immunization with a pentameric L1 fusion protein protects against papillomavirus infection.

The prophylactic papillomavirus vaccines currently in clinical trials are composed of viral L1 capsid protein that is synthesized in eukaryotic expression systems and purified in the form of virus-like particles (VLPs). To evaluate whether VLPs are necessary for effective vaccination, we expressed the L1 protein as a glutathione S-transferase (GST) fusion protein in Escherichia coli and assayed its immunogenic activity in an established canine oral papillomavirus (COPV) model that previously validated the efficacy of VLP vaccines. The GST-COPV L1 fusion protein formed pentamers, but these capsomere-like structures did not assemble into VLPs. Despite the lack of VLP formation, the GST-COPV L1 protein retained its native conformation as determined by reactivity with conformation-specific anti-COPV antibodies. Most importantly, the GST-COPV L1 pentamers completely protected dogs from high-dose viral infection of their oral mucosa. L1 fusion proteins expressed in bacteria represent an economical alternative to VLPs as a human papillomavirus vaccine.

Preclinical safety evaluation of G207, a replication-competent herpes simplex virus type 1, inoculated intraprostatically in mice and nonhuman primates.

G207, a replication-competent herpes simplex virus type 1 (HSV-1) virus, has been previously shown to be effective against human prostate cancer xenografts in mice. This study assesses its safety in the prostate of two animal models known for their sensitivity to HSV-1. BALB/c mice were injected intraprostatically with either HSV-1 G207 or strain F and observed for 5 months. None of the G207-injected animals exhibited any clinical signs of disease or died. However, 50% of strain F-injected mice displayed sluggish, hunched behavior and died by day 13. Histopathologically, the G207-injected prostates were normal whereas strain F-injected prostates showed epithelial flattening, sloughing, and stromal edema. Four Aotus nancymae monkeys were also injected with G207 intraprostatically and observed short term (up to 21 days) and long term (56 days). Safety was assessed on the basis of clinical observations, viral biodistribution, virus shedding, and histopathology. None of the injected monkeys displayed evidence of clinical disease, shedding of infectious virus, or spread of the virus into other organs. Except for minor histological changes unrelated to the study, no significant abnormalities were observed. These results demonstrate that G207 can be safely inoculated into the prostate and should be considered for human trials for the treatment of prostate cancer.

Experimental methodologies and preliminary transfer factor data for estimation of dermal exposures to particles.

Developmental efforts and experimental data that focused on quantifying the transfer of particles on a mass basis from indoor surfaces to human skin are described. Methods that utilized a common fluorescein-tagged Arizona Test Dust (ATD) as a possible surrogate for housedust and a uniform surface dust deposition chamber to permit estimation of particle mass transfer for selected dust size fractions were developed. Particle transfers to both wet and dry skin were quantified for contact events with stainless steel, vinyl, and carpeted surfaces that had been pre-loaded with the tagged test dust. To better understand the representativeness of the test dust, a large housedust sample was collected and analyzed for particle size distribution by mass and several metals (Pb, Mn, Cd, Cr, and Ni). The real housedust sample was found to have multimodal size distributions (mg/g) for particle-phase metals. The fluorescein tagging provided surface coatings of 0.11-0.36 ng fluorescein per gram of dust. The predominant surface location of the fluorescein tag would best represent simulated mass transfers for contaminant species coating the surfaces of the particles. The computer-controlled surface deposition chamber provided acceptably uniform surface coatings with known particle loadings on the contact test panels. Significant findings for the dermal transfer factor data were: (a) only about 1/3 of the projected hand surface typically came in contact with the smooth test surfaces during a press; (b) the fraction of particles transferred to the skin decreased as the surface roughness increased, with carpeting transfer coefficients averaging only 1/10 those of stainless steel; (c) hand dampness significantly increased the particle mass transfer; (d) consecutive presses decreased the particle transfer by a factor of 3 as the skin surface became loaded, requiring approximately 100 presses to reach an equilibrium transfer rate; and (e) an increase in metals concentration with decreasing particle size, with levels at 25 microns typically two or more times higher than those at 100 microns--consistent with the earlier finding of Lewis et al. for the same sample for pesticides and polycyclic aromatic hydrocarbons (PAHs).

Viral shedding and biodistribution of G207, a multimutated, conditionally replicating herpes simplex virus type 1, after intracerebral inoculation in aotus.

G207 is a multimutated, conditionally replicating herpes simplex virus type 1 (HSV-1) that is currently in clinical trial for patients with malignant glioma. G207 exhibits an efficient oncolytic activity in tumor cells, yet minimal toxicity in normal tissue when injected into the brains of HSV-susceptible mice or nonhuman primates. In this study, we evaluated the shedding and biodistribution of clinical-grade G207 after intracerebral inoculation (3 x 10(7) pfu) in four New World owl monkeys (Aotus nancymae). Using PCR analyses and viral cultures, neither infectious virus nor viral DNA was detected from tear, saliva, or vaginal secretion samples at any time point up to 1 month postinoculation. Analyses of tissues obtained at necropsy at 1 month from two of the four monkeys, plus one monkey inoculated with laboratory-grade G207 (10(9) pfu) 2 years earlier, showed the distribution of G207 DNA restricted to the brain, although infectious virus was not isolated. Histopathology revealed normal brain tissues including the sites of inoculation. A measurable increase of serum anti-HSV antibody titer was observed in all monkeys, as early as 21 days postinoculation. The results ascertain the safety of G207 in the brain and indicate that strict biohazard management may not be required for G207-treated patients.

The effect of sodium bicarbonate on propranolol-induced cardiovascular toxicity in a canine model.

STUDY OBJECTIVE: To evaluate the potential utility of sodium bicarbonate in an established model of acute propranolol toxicity. METHODS: Two minutes after the completion of a propranolol infusion (10 mg/kg), a bolus of 1.5 mEq/kg of sodium bicarbonate solution (1 mEq/mL) followed by an infusion of 1.5 mEq/kg over the next 26 minutes (n = 6) or an equivalent timing and volume of 5% dextrose solution (n = 6) was administered in each dog. Targeted cardiovascular parameters included heart rate, mean arterial pressure, left ventricular dP/dtmax, and QRS interval. RESULTS: Propranolol infusion significantly depressed heart rate (p < 0.0001), mean arterial pressure (p < 0.0001), dP/dtmax (p < 0.0001) and prolonged the QRS interval (p < 0.0001). Sodium bicarbonate failed to significantly improve these targeted parameters when compared to control animals. CONCLUSION: In this canine model of propranolol toxicity, intravenous sodium bicarbonate appears to be an ineffective single therapy. Furthermore, these results may suggest a different mechanism of sodium channel blockade for propanolol than that of type IA antiarrhythmic agents.

A phase I study of combined UFT plus leucovorin and radiotherapy for pancreatic cancer.

PURPOSE: This Phase I study combines tegafur and uracil (UFT) with leucovorin and conventional radiation for the treatment of pancreatic cancer. The design seeks to determine the maximum tolerated dose (MTD) and dose-limiting toxicities (DLT) of this regimen as well as to define a future Phase II dose level. METHODS: Patients with locally advanced and unresectable pancreatic cancer were treated with 45 Gy of radiation therapy. The initial UFT dose was 150 mg/m(2)/day given with leucovorin 90 mg/day, both divided into 3 daily doses for 35 days concurrent with radiation. UFT doses were escalated at increments of 50 mg/m(2)/day. Dose-limiting toxicity (DLT) was defined as Grade 3 or greater nausea, vomiting or diarrhea despite medical intervention; or Grade 3 or greater neutropenia/thrombocytopenia; or Grade 3 or greater hepatic toxicity; or inability of the patient to take 75% or more of the planned UFT/leucovorin; or radiotherapy interruption of greater than 1 week. The MTD for UFT/leucovorin was exceeded by one dose level when a certain dose caused DLT in 2 or more patients of 6. RESULTS: Five evaluable patients had Stage I resectable disease but had pathologic adenopathy. Seven had Stage II unresectable disease. Compliance with therapy was excellent. At a daily dose of 300 mg/m(2) of UFT, we noticed minimal diarrhea and hematologic toxicity with mild-moderate nausea, anorexia, and fatigue. Three patients had Grade 4 toxicity: 1 had neutropenia on Day 38, 1 had diarrhea on Day 55, and 1 had vomiting on Day 15. CONCLUSION: Oral UFT/leucovorin and radiation therapy offers patients a viable treatment option for pancreatic cancer. The major known toxicity of diarrhea was tolerable. The MTD was not reached in this study. Our current plan is to expand this into a Phase I/II trial beginning at a UFT dose of 300 mg/m(2) and correlate this with clinical pharmacologic parameters. The potential benefit of long bioavailability and oral delivery of UFT compares favorably with continuous infusion regimens without the added morbidity of a catheter and pump.

Spontaneously regressing oral papillomas induce systemic antibodies that neutralize canine oral papillomavirus.

Canine oral papillomavirus (COPV) infection of naive beagle dogs causes oral papillomas, most of which spontaneously regress. Regressor beagles do not develop new oral papillomas because of COPV type-specific, cell-mediated immunity, COPV neutralizing antibodies, or both. Formalin-fixed native and recombinant COPV vaccines that target the systemic immune system induce neutralizing antibodies that prevent development of oral papillomas. This study was designed to determine whether spontaneously regressing mucosal papillomas also targeted the systemic immune system to induce circulating, neutralizing IgG antibodies that protect against infection by COPV. To accomplish this goal, IgG was fractionated from sera collected from weanling beagles and regressor beagles and tested for conferring protection by passive immunization. Serum was tested by ELISA for antibodies against intact virions and then pooled for passive transfer to naive beagles. Preimmune sera were neither reactive by ELISA nor protective by passive transfer. On the other hand, IgG antibodies from regressor beagles were reactive by ELISA and passive transfer conferred protection against COPV challenge. Circulating IgG antibodies induced by spontaneous regression of canine oral papillomas protect beagles against intraoral infection by COPV, a model for mucosotropic HPV.

Attenuated, replication-competent herpes simplex virus type 1 mutant G207: safety evaluation of intracerebral injection in nonhuman primates.

This study examined the safety of intracerebral inoculation of G207, an attenuated, replication-competent herpes simplex virus type 1 (HSV-1) recombinant, in nonhuman primates. Sixteen New World owl monkeys (Aotus nancymae [karyotype 1, formerly believed to be A. trivirgatus]), known for their exquisite susceptibility to HSV-1 infection, were evaluated. Thirteen underwent intracerebral inoculation with G207 at doses of 10(7) or 10(9) PFU, two were vehicle inoculated, and one served as an infected wild-type control and received 10(3) PFU of HSV-1 strain F. HSV-1 strain F caused rapid mortality and symptoms consistent with HSV encephalitis, including fever, hemiparesis, meningitis, and hemorrhage in the basal ganglia. One year after G207 inoculation, seven of the animals were alive and exhibited no evidence of clinical complications. Three deaths resulted from nonneurologic causes unrelated to HSV infection, and three animals were sacrificed for histopathologic examination. Two animals were reinoculated with G207 (10(7) PFU) at the same stereotactic coordinates 1 year after the initial G207 inoculation. These animals were alive and healthy 2 years after the second inoculation. Cerebral magnetic resonance imaging studies performed both before and after G207 inoculation failed to reveal radiographic evidence of HSV-related sequelae. Despite the lack of outwardly observable HSV pathology, measurable increases in serum anti-HSV titers were detected. Histopathological examination of multiple organ tissues found no evidence of HSV-induced histopathology or dissemination. We conclude that intracerebral inoculation of up to 10(9) PFU of G207, well above the efficacious dose in mouse tumor studies, is safe and therefore appropriate for human clinical trials.

Control of negative inotropic vagal preganglionic neurons in the dog: synaptic interactions with substance P afferent terminals in the nucleus ambiguus?

Previous research from this laboratory has shown that substance P-immunoreactive (SP) terminals synapse upon negative chronotropic vagal preganglionic neurons (VPNs), but not upon negative dromotropic VPNs, of the ventrolateral nucleus ambiguus (NA-VL). Moreover, SP agonists injected into NA-VL cause bradycardia without decreasing AV conduction. In the current study, we have: (1) defined the electron microscopic characteristics of the SP neurons of NA-VL in dog; and (2) tested the hypothesis that SP nerve terminals synapse upon negative inotropic VPNs of NA-VL, retrogradely labeled from the cranial medial ventricular (CMV) ganglion. Numerous SP terminals and a few SP neurons were observed in the vicinity of retrogradely labeled neurons. SP terminals were observed forming synapses with unlabeled dendrites and with SP dendrites, but never with the retrogradely labeled neurons. Together, these results and earlier findings suggest that SP agonists may be able to induce bradycardia without decreasing AV conduction or ventricular contractility.

Up-regulation of two Candida albicans genes in the rat model of oral candidiasis detected by differential display.

Candida albicans is an opportunistic fungal pathogen responsible for the largest percentage of fungal-mediated oral and oesophageal disease. In this regard, knowledge concerning patterns of gene expression during the establishment and/or maintenance of infection may be the key to the design of new strategies for treatment, as well as providing insight into pathogenesis. To address this issue, experiments were performed that utilized differential display to compare the spectrum of C. albicans genes expressed during oral infection versus growth in in vitroculture. Experimentally, the rat model of oral candidiasis served as the in vivo source. After initiation of infection and subsequent harvesting of C. albicans from the rat oral cavity, RNA was isolated, and used with a small number of primers in reverse-transcriptase polymerase chain reaction (RT-PCR) and differential display experiments. Fragments unique to in vivo samples were subcloned and sequenced. Southern blot analysis verified the origin of seven fragments as fromC. albicans. Additionally, specific RT-PCR confirmed that two of these fragments represented genes that were up-regulated during C. albicans in vivo growth in the rat model. Database searches indicated the fragments share homology with a member of the C. albicans agglutinin gene family and to a bacterial gene (gidB) possibly involved in cell division.

Neural control of left ventricular contractility in the dog heart: synaptic interactions of negative inotropic vagal preganglionic neurons in the nucleus ambiguus with tyrosine hydroxylase immunoreactive terminals.

Recent physiological evidence indicates that vagal postganglionic control of left ventricular contractility is mediated by neurons found in a ventricular epicardial fat pad ganglion. In the dog this region has been referred to as the cranial medial ventricular (CMV) ganglion [J.L. Ardell, Structure and function of mammalian intrinsic cardiac neurons, in: J.A. Armour, J.L. Ardell (Eds.). Neurocardiology, Oxford Univ. Press, New York, 1994, pp. 95-114; B.X. Yuan, J.L. Ardell, D.A. Hopkins, A.M. Losier, J.A. Armour, Gross and microscopic anatomy of the canine intrinsic cardiac nervous system, Anat. Rec., 239 (1994) 75-87]. Since activation of the vagal neuronal input to the CMV ganglion reduces left ventricular contractility without influencing cardiac rate or AV conduction, this ganglion contains a functionally selective pool of negative inotropic parasympathetic postganglionic neurons. In the present report we have defined the light microscopic distribution of preganglionic negative inotropic neurons in the CNS which are retrogradely labeled from the CMV ganglion. Some tissues were also processed for the simultaneous immunocytochemical visualization of tyrosine hydroxylase (TH: a marker for catecholaminergic neurons) and examined with both light microscopic and electron microscopic methods. Histochemically visualized neurons were observed in a long slender column in the ventrolateral nucleus ambiguus (NA-VL). The greatest number of retrogradely labeled neurons were observed just rostral to the level of the area postrema. TH perikarya and dendrites were commonly observed interspersed with vagal motoneurons in the NA-VL. TH nerve terminals formed axo-dendritic synapses upon negative inotropic vagal motoneurons, however the origin of these terminals remains to be determined. We conclude that synaptic interactions exist which would permit the parasympathetic preganglionic vagal control of left ventricular contractility to be modulated monosynaptically by catecholaminergic afferents to the NA-VL.

Gamma interferon prevents diabetes in the BB rat.

The BB rat model of human insulin-dependent diabetes mellitus (IDDM) spontaneously develops diabetes through an autoimmune process. Gamma interferon (IFN-gamma) is thought to play an important pathogenic role. This study examined if IFN-gamma administration can, paradoxically, prevent diabetes in BB rats. Diabetes-prone BB rats were initially injected intraperitoneally with murine recombinant IFN-gamma (rIFN-gamma) at doses of 0.5 x 10(4) to 40 x 10(4) U three times a week for 6 weeks beginning at 35 days of age. The effects of altering the duration of treatment (2 to 6 weeks) and the age at which injections were initiated (45 through 65 days) were also assessed. rIFN-gamma administration prevented the development of diabetes in a dose-dependent manner. The optimal treatment condition resulted in a 9.1% incidence of diabetes versus a 90% incidence in control rats. This diabetes-sparing effect was long lasting and continued to 7 months of age. A 4- to 6-week course resulted in maximal inhibition. Treatment initiated as late as 55 days of age, when insulitis is already present, was effective in preventing diabetes. Islet inflammation was dramatically lower in rIFN-gamma- versus saline-injected rats (P < 0.01). Total leukocyte count and subpopulations of peripheral mononuclear cells were unaltered by rIFN-gamma. In summary, rIFN-gamma paradoxically and potently prevents diabetes in BB rats in a dose-dependent fashion by inhibiting islet inflammation. This diabetes-sparing effect occurs even when injections are initiated after evidence of the diabetic process is already present.

Paired comparison of vascular wall reactions to Palmaz stents, Strecker tantalum stents, and Wallstents in canine iliac and femoral arteries.

BACKGROUND: Palmaz stents, Strecker stents, and Wallstents, all used clinically, differ substantially in their physical characteristics, yet how differently the vascular wall reacts to them has not been demonstrated conclusively. We therefore undertook a side-by-side comparison. METHODS AND RESULTS: One stent was implanted into each canine external iliac and/or the flexing portion of the proximal femoral artery. In 9 dogs, Palmaz stents were placed vis-à-vis Strecker stents, with follow-up of 2 and 4 months. In 7 dogs, Palmaz stents were placed vis-à-vis Wallstents, with 4 months of follow-up. Angiographic midstent luminal diameters immediately after placement and at follow-up as well as midstent cross-sectional areas of neointima were compared for significant differences. In addition, neointimal maturation, medial atrophy, and stent-related trauma were assessed. Angiographically, all arteries remained open. The degree of luminal narrowing by recoil and neointima never reached 50% and was modest for Palmaz stents and Wallstents (P = .33) but significantly higher for Strecker stents (P < .0001 compared with Palmaz stents). This corresponded histologically to a significantly thicker neointima (P = .003) over Strecker than over Palmaz stents but not between Palmaz stents and Wallstents (P = .18). Neointimal buildup was generally more pronounced in the femoral artery segments than in the iliac segments. Maturation of the neointima over Palmaz stents was much further advanced than over Strecker stents and slightly more advanced than over Wallstents. Pressure-related atrophy of the tunica media was least for Strecker stents and more pronounced but similar for Wallstents and Palmaz stents. Wallstent wire ends caused some wall trauma; several femoral Palmaz stent struts protruded through the media. CONCLUSIONS: The lower-hoop-strength, higher-profile tantalum Strecker stent is affected by vascular wall recoil and evokes a greater degree of neointima formation than the lower-profile, higher-hoop-strength Palmaz stent and Wallstent. Medial atrophy is pronounced outside the latter two stents. The rigid Palmaz stent can penetrate through the vascular wall in flexing arteries.

Systemic immunization with papillomavirus L1 protein completely prevents the development of viral mucosal papillomas.

Infection of mucosal epithelium by papillomaviruses is responsible for the induction of genital and oral warts and plays a critical role in the development of human cervical and oropharyngeal cancer. We have employed a canine model to develop a systemic vaccine that completely protects against experimentally induced oral mucosal papillomas. The major capsid protein, L1, of canine oral papillomavirus (COPV) was expressed in Sf9 insect cells in native conformation. L1 protein, which self-assembled into virus-like particles, was purified on CsCl gradients and injected intradermally into the foot pad of beagles. Vaccinated animals developed circulating antibodies against COPV and became completely resistant to experimental challenge with COPV. Successful immunization was strictly dependent upon native L1 protein conformation and L1 type. Partial protection was achieved with as little as 0.125 ng of L1 protein, and adjuvants appeared useful for prolonging the host immune response. Serum immunoglobulins passively transferred from COPV L1-immunized beagles to naive beagles conferred protection from experimental infection with COPV. Our results indicate the feasibility of developing a human vaccine to prevent mucosal papillomas, which can progress to malignancy.

Host response to multiple skin lacerations on the albino guinea pig.

Investigators have studied methods of treating skin lacerations by placing multiple incisions on each albino guinea pig. Theoretically, host responses to laceration sites may differ on the basis of anatomic location and local cytokine effects. We used cytokine values and histologic examination to identify differences when multiple lacerations were placed on each animal. Four 3-cm lacerations were made on the dorsum of each male albino guinea pig: two incisions on either side and parallel to the spine. Each laceration was closed with staples. In five animals a sponge technique was used to assay wound cytokines 48 h later. In an additional four animals, wounds were excised at 96 h and stained for cells and new collagen. We identified no statistically significant differences among laceration sites based on polymorphonuclear and mononuclear cellularity, number of fibroblasts, new collagen deposition, or wound interleukin (IL)-6 activities. Using this model minimizes the number of animals needed to generate statistically significant findings in wound research.