Immunohistochemical localization of renin in mouse brain

Immunohistochemical studies of mouse brain by the peroxidase-antiperoxidase method using monospecific antimouse renin antibodies has revealed the intracellular localization of renin in stellate and small ovoid cells. Renin-containing ovoid cells were observed in the spinal cord, medulla oblongata, pons, granular layers of the cerebellum, deep cerebellar region, and the lamina terminalis; whereas immunostainable stellate cells were found in the cerebral cortex, hippocampus, dentate gyrus and septum. Intracellular localization of renin rather than intravascular localization supports an endogenous origin of this enzyme in the brain. Wide distribution in different types of cells suggests different types of regulatory mechanisms. Double immunostaining with antigalactocerebroside and antirenin antibodies indicates the presence of renin in oligodendrocytes.     

Alterations in epithelial glycocalyx of rabbit uteri during early pseudopregnancy and pregnancy,and following ovariectomy

Pseudopregnant, pregnant, and ovariectomized rabbits were utilized to study hormonal mediation of uterine epithelial surface negativity and glycocalyx morphology, and to seek local effects of blastocysts at sites of implantation. A loss of surface negativity [polycationic ferritin (PCF) binding] by day 6 of pregnancy or pseudopregnancy was noted, accompanied by alterations in epithelial glycocalyx. Uteri from estrous animals, or ovariectomized animals receiving oil or estradiol injections, bound PCF and exhibited a “globular” glycocalyx. Uteri from day 6 pseudopregnant or pregnant animals, or ovariectomized animals receiving progesterone injections, did not bind PCF or exhibit a globular glycocalyx. Both PCF binding and the globular character of the epithelial glycocalyx were sensitive to neuraminidase and trypsin treatment, suggesting sialoglycoprotein contribution to surface negativity. Implanting blastocysts had no detectable local effect on surface negativity, but did induce local reduction of epithelial glycocalyx at sites of implantation. Results of this study suggest that uterine epithelial glycocalyx alterations during the preimplantation period reflect a general response to prosgesterone stimulation, primarily qualtitative in nature, related to the acquisition of receptivity to ovo-implantation.     

Survival,growth, and accumulation of ingested lead in nestling American Kestrels (Falco sparverius)

One-day old American Kestrel (Falco sparverius) nestlings were dosed orally daily with 5 l/g of corn oil (controls), 25 mg/kg, 125 mg/kg, or 625 mg/kg of metallic lead in corn oil through day 10. Forty percent of the nestlings given 625 mg/kg died after six days. Growth rates became significantly different from controls in the 625 mg/kg group by day 3 and in the 125 mg/kg group by day 4. Crown-rump lengths and brain weights were significantly lower in both treatment groups. Liver and kidney weights were lower in the 625 mg/kg groups. Skeletal examination and measurement of alizarin red-S stained nestlings revealed reduced growth for the humerus, radius-ulna, femur, and tibiotarsus in the 125 mg/kg and 625 mg/kg groups. Skeletons were otherwise normal in appearance. Greater than 2 ppm (wet weight) lead in the liver or 6 ppm in the kidney was associated with suppressed growth, while more than 5 ppm in the liver and 15 ppm in the kidney occurred in survivors in the 625 mg/kg group. The order of accumulation of lead in tissues at the end of 10 days was kidney > liver > brain. These findings suggest that altricial nestlings may be considerably more sensitive to lead exposure than adults and also more sensitive than hatchlings of many precocial species.     

Survival,growth, and histopathological effects of paraquat ingestion in nestling american kestrels(Falco sparverius)

The use of paraquat as a herbicide is becoming more extensive with the increasing popularity of no tillage agriculture, increasing the possibility of exposure for wildlife species. American kestrel(Falco sparverius) nestlings were orally dosed daily with 5 l/g of distilled water (controls), 10 mg/kg, 25 mg/kg, or 60 mg/kg of paraquat dichloride (1,1-dimethyl-4,4-bipyridinium) in distilled water from day 1 through day 10. Forty-four percent of the nestlings given 60 mg/kg died after 4 days. Significant differences in growth rates occurred between controls and all paraquat-dosed groups. Reduced skeletal growth occurred in the humerus and femur in the 25 mg/kg and 60 mg/kg groups, and in the radius-ulna and tibiotarsus in the 60 mg/kg group. Skeletons were otherwise normal in appearance. Histopathological examination revealed localized focal necrosis in the liver of one nestling in the 60 mg/kg group and tubular cell degeneration and focal tubular dilation in the kidneys of another. The brain and lungs were unremarkable histologically. These findings suggest that altricial nestling kestrels are more sensitive to paraquat exposure than young or adult birds of precocial species.     

Lymphokine-activated killer (LAK) cells can be focused at sites of tumor growth by products of macrophage activation

Successful adoptive cancer immunotherapy presumably depends on the accumulation of tumoricidal leukocytes at the sites of tumor growth. Large numbers of lymphokine-activated killer (LAK) cells can be generated in vitro by growth in high concentrations of interleukin-2 (IL-2), but relatively few arrive at the tumor site after intravenous injection. We hypothesize that the delivery of LAK cells to tumor sites may be augmented by previously demonstrated lymphocyte-recruiting factors, including activated macrophage products such as interleukin-1 (IL-1) and tumor necrosis factor. 111Indium-labeled LAK cells were injected intravenously into syngeneic mice bearing the macrophage activator endotoxin (LPS) in one hind footpad, and saline solution was injected into the contralateral footpad. Significantly more activity was recovered from the LPS-bearing footpad at all times during a 96-hour period. Recombinant IL-1 also attracted more LAK cells after injection into tumor-free hind footpads. Furthermore, LAK cells preferentially homed to hind footpads that were bearing 3-day established sarcomas after intralesional injections of LPS, IL-1, or tumor necrosis factor when compared with contralateral tumor-bearing footpads injected with saline solution alone. In preliminary experiments, mice with hind-footpad tumors appeared to survive longer after combined systemic IL-2 and LAK therapy if intralesional LPS was administered. These studies demonstrate that macrophage activation factors that have been shown capable of attracting circulating normal lymphocytes can also effectively attract LAK cells from the circulation. By the stimulation of macrophages at the sites of tumor growth, more LAK cells can be attracted. It is hoped that by "focusing" the migration of LAK cells to tumors, LAK cells and IL-2 would effect tumor regression more efficiently and with less toxicity.     

The impact of SARS-CoV-2 on the cystic fibrosis foundation therapeutics development network

The Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV-2) global pandemic significantly impacted CF clinical research within the Cystic Fibrosis Foundation Therapeutics Development Network (CFF TDN). A Research Electronic Data Capture (REDCap) survey was developed and sent to network sites to monitor and understand the impact on research teams, ongoing and anticipated clinical research, and specific clinical and research procedures. Key findings indicated an early impact on participant enrollment, research team stability, and procedures such as spirometry and sputum induction. These trends steadily improved over the months as research activities began to recover across the TDN. While SARS-CoV-2 created a significant challenge it also highlights new opportunities to expand CF research with greater focus on data collection outside of research centers and increased access for remote participation.     

A randomized controlled trial evaluating combination detection of HIV in Malawian sexually transmitted infections clinics

IntroductionHIV diagnosis is the necessary first step towards HIV care initiation, yet many persons living with HIV (PLWH) remain undiagnosed. Employing multiple HIV testing strategies in tandem could increase HIV detection and promote linkage to care. We aimed to assess an intervention to improve HIV detection within socio‐sexual networks of PLWH in two sexually transmitted infections (STI) clinics in Lilongwe, Malawi.MethodsWe conducted a randomized controlled trial to evaluate an intervention combining acute HIV infection (AHI) screening, contract partner notification and social contact referral versus the Malawian standard of care: serial rapid serological HIV tests and passive partner referral. Enrolment occurred between 2015 and 2019. HIV‐seropositive persons (two positive rapid tests) were randomized to the trial arms and HIV‐seronegative (one negative rapid test) and ‐serodiscordant (one positive test followed by a negative confirmatory test) persons were screened for AHI with HIV RNA testing. Those found to have AHI were offered enrolment into the intervention arm. Our primary outcome of interest was the number of new HIV diagnoses made per index participant within participants’ sexual and social networks. We also calculated total persons, sexual partners and PLWH (including those previously diagnosed) referred per index participant.ResultsA total of 1230 HIV‐seropositive persons were randomized to the control arm, and 561 to the intervention arm. Another 12,713 HIV‐seronegative or ‐serodiscordant persons underwent AHI screening, resulting in 136 AHI cases, of whom 94 enrolled into the intervention arm. The intervention increased the number of new HIV diagnoses made per index participant versus the control (ratio: 1.9; 95% confidence interval (CI): 1.2 to 3.1). The intervention also increased the numbers of persons (ratio: 2.5; 95% CI: 2.0 to 3.2), sexual partners (ratio: 1.7; 95% CI: 1.4 to 2.0) and PLWH (ratio: 2.3; 95% CI: 1.7 to 3.2) referred per index participant.ConclusionsCombining three distinct HIV testing and referral strategies increased the detection of previously undiagnosed HIV infections within the socio‐sexual networks of PLWH seeking STI care. Combination HIV detection strategies that leverage AHI screening and socio‐sexual contact networks offer a novel and efficacious approach to increasing HIV status awareness.     

Accumulation and effects of lead and cadmium on wood ducks near a mining and smelting complex in Idaho

A study of wood ducks (Aix sponsa) was conducted along the Coeur d'Alene River system in northern Idaho in 1986 and 1987. Most of this area has been subjected to severe contamination from lead and other metals from mining and smelting since the 1880s. In 1986, a preliminary study of wood duck nesting was conducted in the contaminated area; incubating hens captured in nest boxes were bled and weighed. Blood samples were used to determine lead and cadmium concentrations and physiological characteristics. In 1987, an intensive study of wood ducks involved trapping and monitoring nest boxes in the contaminated area. Blood and tissue samples were also taken from wood ducks from a reference area without known contamination from metals. Lead levels in blood and tissues of most wood ducks from the contaminated area frequently exceeded those considered hazardous to birds; maximum levels (wet weight) of lead were 8 g g^–1 in blood and 14 g g^–1 in liver. Changes in physiological characteristics constituted the only evidence of potentially adverse effects from lead. In the contaminated area, nesting success (55% unadjusted, 35% Mayfield estimate) was less than in other areas where predation was low and nest boxes were used; but lead concentrations and physiological characteristics of blood were similar in successful and unsuccessful hens.Values of ALAD, hemoglobin, and body mass were negatively correlated with blood concentrations of lead, whereas protoporphyrin was positively correlated with lead levels in the blood. Some of the protoporphyrin values (1,091 g dl^–1 in a male and 756 g dl^–1 in a female) equalled those associated with lead toxicosis in experimental birds. ALAD activity was low in most birds from the contaminated area; values of 0 were obtained from 11 birds. Lead levels in blood, ALAD, protoporphyrin, and hemoglobin were significantly different between birds from the contaminated and reference areas. Concentrations of lead in ingesta of wood ducks ranged from 0.9 to 610 g g^–1 in the contaminated area and 0.2 to 0.6 g g^–1 in the reference area. Levels of cadmium in kidneys of wood ducks ranged from 1g g^–1 to 20 g g^–1 in the contaminated area and from only to 0.1 g g^–1 to 1 g g^–1 in the reference area. Cadmium concentrations were less than known effect levels.