A scoping review of unintended harm associated with public health interventions: towards a typology and an understanding of underlying factors.

OBJECTIVES: Unintended harm theory as related to public health interventions (PHI) is under developed, with harm evaluation and reporting often absent or incomplete. This review presents a typology for, and underlying factors linked to, PHI-associated unintended harm. METHODS: This scoping review was conducted electronically and includes articles from 1992 to June of 2013. Out of 2,490 originally identified titles, 26 full-text articles were included that discussed unintended harm associated with PHI. An iterative data analysis process was utilized to identify both a typology and underlying factors associated with unintended harm. RESULTS: A typology of PHI-associated unintended harm was identified: (1) physical; (2) psychosocial; (3) economic; (4) cultural and (5) environmental. Five underlying factors associated with PHI unintended harm emerged: (1) limited and/or poor quality evidence; (2) prevention of one extreme leads to another (boomerang effects); (3) lack of community engagement; (4) ignoring root causes; and (5) higher-income country PHI implementation in a lower- or middle-income country. CONCLUSIONS: PHI planning and evaluation frameworks may benefit from the consideration and potential incorporation of the unintended harm typology and underlying factors.

Promoting health, preserving culture: adapting RARE in the Maasai context of Northern Tanzania.

HIV/AIDS prevention strategies often neglect traditions and cultural practices relevant to the spread of HIV. The role of women in the HIV/AIDS context has typically been relegated to high-risk female groups such as sex workers, or those engaged in transactional sex for survival. Consequently, these perceptions are born out in the escalation of HIV/AIDS among communities, and female populations in particular where prevention frameworks remain culturally intolerant. We have attempted to address these issues by using an adapted Rapid Assessment Response and Evaluation (RARE) model to examine the impact of HIV/AIDS in the Maasai community of Ngorongoro. Our adapted RARE model used community engagement venues such as stockholder workshops, key informant interviews, and focus groups. Direct observations and geomapping were also done. Throughout our analysis, a gender and a pastoralist-centered approach provided methodological guidance, and served as value added contributions to out adaptation. Based in the unique context of a rural pastoralist community, we made recommendations appropriate to the cultural setting and the RARE considerations.

The influence of basic fibroblast growth factor on cat corneal endothelial wound healing in vivo.

Using standardized freeze wounds in cat corneas, we tested the efficacy of basic Fibroblast Growth Factor (bFGF) solubilized in phosphate buffered saline (PBS) to promote endothelial healing when injected intraocularly at doses ranging from 0.01 microgram to 10 micrograms. After 6 days, animals were humanely sacrificed and corneal tissues were fixed and stained for light microscopy and computation of remaining wound areas. A significant dose-response relationship was found between the dosages of 0.01, 0.1, and 1.0 microgram of bFGF/eye and the stimulation of more completely healed endothelium six days after transcorneal freeze wounding. Significantly larger endothelial wounds were present six days after wounding when the eyes were treated with 10 micrograms of bFGF/eye compared with controls treated with PBS only.

The influence of epidermal growth factor on cat corneal endothelial wound healing.

Using standardized freeze wounds in cat corneas, we tested the efficacy of human recombinant Epidermal Growth Factor (EGF) to promote endothelial healing when solubilized in either phosphate buffered saline (PBS), 1% methylcellulose (MC), or sodium hyaluronate (NaHA), in final intraocular doses ranging from 2 micrograms to 100 micrograms of EGF. After 6 or 7 days' healing, animals were humanely sacrificed and corneal tissues were fixed and stained for light microscopy and computation of remaining wound areas. EGF in NaHA in final intraocular doses of 2 and 10 micrograms prompted significantly more complete healing of transcorneal freeze wounds to endothelium compared with endothelium of eyes treated with NaHA control solution alone. EGF in PBS or in MC in doses ranging from 2-100 micrograms/eye did not promote more complete wound healing than that seen in eyes treated with their respective vehicle solutions alone. All vehicle solutions were associated with similar degrees of wound healing, implying that they have no intrinsic healing properties.

Stimulation of corneal endothelial wound healing in cats by mesodermal growth factor.

Several growth factors have been evaluated for their effects on corneal wound healing but few studies have yet demonstrated an acceleration of endothelial repair in vivo. Mesodermal Growth Factor (MGF) was tested in vivo by making standardized freeze wounds in cat corneas and immediately injecting one of four concentrations of MGF in sterile phosphate buffered saline (PBS), or PBS alone, into the anterior chamber. Seven days later, the animals were sacrificed and the corneas excised and stained. Descemet's membrane and endothelium were dissected and mounted onto glass slides. The wound areas were photographed, measured and compared statistically. Those cats receiving the three lowest doses of MGF had significantly smaller wounds than controls (p less than 0.05).

In situ hybridization detection of marked differences in pre-proopiomelanocortin messenger ribonucleic acid content of individual corticotropes and melanotropes.

Recently, heterogeneity of POMC mRNA content between intermediate lobe melanotropes of the rat pituitary gland was demonstrated by in situ hybridization of tissue sections. In the present study the heterogeneity of POMC mRNA content in dispersed rat pituitary cells has been investigated. Acutely dispersed cells from adult male rat anterior or neurointermediate lobe tissues were adhered to poly-L-lysine-coated coverslips. The cells were fixed and then hybridized with 35S-labeled POMC or 1B15 (cyclophilin) cRNA. Parallel studies measuring constitutively expressed cellular 1B15 mRNA content were undertaken to ensure that the apparent single cell differences in POMC mRNA were not inherent to the in situ hybridization procedure. When classified by image analysis, extensive differences in silver grain densities were seen over POMC mRNA-containing cells from both lobes. To determine if mRNA in polysomal configurations was less accessable for hybridization with probes than naked mRNA, cells were preincubated with pactamycin, a potent inhibitor of ribosomal initiation of protein synthesis. Pactamycin had no effect on these results. Thus, there appears to be large differences in POMC mRNA content between individual pituitary cells expressing the same gene product.

Post-translational processing of pro-opiomelanocortin (POMC)-derived peptides during fetal monkey pituitary development. II. beta-Lipotropin (beta-LPH)-related peptides.

We have studied the post-translational processing of POMC-derived peptides during fetal monkey pituitary development using immunoassay and reverse-phase high-performance liquid chromatography (RP HPLC). Whole pituitary glands obtained from Day 50 and 55 fetal monkeys and separated lobes From Day 65 to 155 were extracted, fractionated, and analyzed for beta-melanotropin (beta-MSH), midportion beta-endorphin (beta-EP), and acetylated beta-EP immunoactivity. Separated adult pituitary lobes were analyzed for comparison. At Day 50, POMC-containing cells were located in both the anterior and intermediate pituitary lobes by immunofluorescence staining, the majority of these cells were localized in the anterior lobe. The Day 50 and 55 whole pituitaries contained predominantly beta-lipotropin (beta-LPH), gamma-lipotropin (gamma-LPH), beta-EP(1-31), and 2.2-kda beta-MSH. No acetylated products were found in Day 50 whole pituitary extracts. By Day 55, carboxy-shortened and acetylated beta-EPs were barely detectable in whole pituitary extracts. These forms were more apparent in the Day 65 separated neurointermediate lobe (NIL) extracts, and were similar to adult proportions by Day 80. The adult anterior lobe contained predominantly beta-LPH, beta-EP, and gamma-LPH. Adult NILs contained almost exclusively 2.2-kda beta-MSH, alpha-N-acetyl beta-EP(1-31) and alpha-N-acetyl beta-EP(1-27). The production of 2.2-kda beta-LPH in the monkey NIL indicates that monkey beta-LPH is different from rat beta-LPH in that it must contain the paired-basic cleavage site required for the formation of 2.2-kda beta-MSH that is known to be lacking in rat beta-LPH. Another finding was that monkey beta-EP contains a Tyr residue at position 27 as found in human beta-EP but appears to have the rat Gln substitution at position 31. The post-translational processing patterns characteristic of each lobe were well established by midterm fetal development (Day 80).

Post-translational processing of pro-opiomelanocortin (POMC)-derived peptides during fetal monkey pituitary development. I. Adrenocorticotropin (ACTH) and alpha-melanotropins (alpha-MSHs).

We have studied the post-translational processing of POMC-derived peptides during fetal monkey development using immunoassay and reverse-phase high-performance liquid chromatography (RP HPLC). Pituitary tissues obtained from fetal monkeys ranging from Gestational Day 50 to 155 were fractionated and analyzed for ACTH- and alpha-MSH-related peptides and compared to adult forms. Extracts of whole pituitary from Fetal Days 50 and 55 contained ACTH(1-39) and very small amounts of CLIP (corticotropin-like intermediate-lobe peptide; ACTH(18-39))-like immunoactivity. Acetylated alpha-MSHs were not detectable at Day 50. alpha-MSHs were barely detectable at Day 55. By Day 65, when pituitary lobes were separable, small amounts of des-, mono-, and diacetyl alpha-MSH were detectable in NIL extracts, but not in anterior lobe extracts. ACTH(1-39) levels were negligible when compared to increasing alpha-MSHs through Fetal Day 80 to 155 in the intermediate lobe. The CLIP immunoactivity was negligible in Day 80 and adult anterior lobe extracts. Thus, lobe-specific proteolytic processing of ACTH-related peptides was well established by midterm gestation. Marked increases of alpha-N- and alpha-N,O-acetylated forms of alpha-MSHs were detected during middle and late stage fetal development. Diacetyl alpha-MSH was the predominant form of alpha-MSH in adult NIL extracts. No acetylated alpha-MSHs were found in anterior lobe tissues, thus adult anterior lobe extracts contained almost exclusively ACTH(1-39). However adult NIL extracts contained two distinct forms of CLIP-related immunoactivity. Therefore changes in post-translational processing patterns of ACTH-related and alpha-MSH-related peptides continued to some extent, postnatally. These data indicate that marked changes in post-translational processing of POMC-derived ACTH-related products occur during the first half of monkey gestation.

Cell bioprocessing in space: applications of analytical cytology.

Cell bioprocessing in space consists of the preparation, cultivation, purification and investigation of cells and their products in the microgravity environment of orbital space flight. Inertial acceleration is used as an independent variable to explore the limits of specific bioprocessing functions, such as cell growth and secretion, gravity-dependent phenomena in cell bioreactors, cell fusion, the influence of thermal convection on processes at cellular dimensions, the electrophoretic separation of cell subpopulations and subcellular particles, and two-phase partitioning of cells, bioparticles, and macromolecules. Analytical cytology techniques are under development for on-orbit application to future cell growth and separation experiments, such as those anticipated in the Space Station era.

Continuous flow electrophoretic separation of proteins and cells from mammalian tissues.

A new continuous flow electrophoretic separator for cells and macromolecules was built and tested in laboratory experiments and in the microgravity environment of space flight. Buffer flows upward in a 120-cm long flow chamber, which is 6 cm wide X 1.5 mm thick in the laboratory version and 16 cm wide X 3.0 mm thick in the microgravity version. Electrophoretic subpopulations are collected in 197 fractions spanning 16 cm at the upper end of the chamber. The electrode buffer is recirculated through front and back cooling chambers, which are also electrode chambers. Ovalbumin and rat serum albumin were used as test proteins in resolution and throughout tests; resolution of these two proteins at 25% total w/v concentration in microgravity was the same as that found at 0.2% w/v concentration in the laboratory. Band spreading caused by Poiseuille flow and conductance gaps was evaluated using polystyrene microspheres in microgravity, and these phenomena were quantitatively the same in microgravity as in the laboratory. Rat anterior pituitary cells were separated into subpopulations enriched with cells that secrete specific hormones; growth-hormone-secreting cells were found to have high electrophoretic mobility, whereas prolactin-secreting cells were found to have low electrophoretic mobility. Cultured human embryonic kidney cells were separated into several electrophoretic subfractions that produced different plasminogen activators; a medium-high-mobility subpopulation and a medium-low-mobility subpopulation each produced a different molecular form of urokinase, whereas a high- and an intermediate-mobility subpopulation produced tissue plasminogen activator. Canine pancreatic islets of Langerhans cells were separated into subpopulations, which, after reaggregation into pseudoislets, were found to be enriched with cells that secrete specific hormones; insulin-secreting beta cells were found in lowest mobility fractions, whereas glucagon-secreting alpha cells were found in the highest mobility fractions. Results of particle electrophoresis experiments were comparable in microgravity and in the laboratory, since cell densities that overloaded the carrier buffer (resulting in zone sedimentation) were avoided, and a 500-fold increase in protein throughput was achieved without compromising resolution in microgravity.

Flow cytometric analysis and sorting of live male rat anterior pituitary cell types by forward angle and perpendicular light scatter.

Light scatter patterns produced by living cells in the flow cytometer are known to provide useful information with regard to their size and internal structure. The purpose of this study was to determine if light scatter signals produced by live male rat anterior pituitary cells could be used as markers to aid in the identification and separation of different hormone-containing cell types. The typical light scatter pattern (4 X 10(5) cells/sample X 15 min) had three ridges in the forward angle light scatter (FALS) perpendicular light scatter (PLS) bivariate cell distribution. FALS signals could be correlated with the size of different cell types and PLS signals with their content of cytoplasmic secretory granules. Agranular cells dominated the low PLS ridge while moderately granulated PRL cells and heavily granulated GH cells dominated the medium and high PLS ridges, respectively. These light scatter patterns were reproducible both within and between different cell suspensions. Inclusion of dopamine in the pituitary gland dissociation medium, a treatment known to increase intracellular PRL contents of mammotrophs, increased the intensity of the PLS signals of a large population of cells, presumably PRL cells. Pituitary cells prepared from different aged male rats also showed changes in light scatter. Cell sorting on the basis of FALS-PLS signals established the relationship between cell type and light scatter pattern.

Flow cytometric analysis and sorting of live female rat anterior pituitary cell types by forward angle and perpendicular light scatter: effect of 17 beta-estradiol.

Administration of physiological concentrations of 17 beta-estradiol (E2) to ovariectomized rats resulted in changes in the forward angle light scatter (FALS) and perpendicular light scatter (PLS) signals of anterior pituitary lobe cells. One day after steroid treatment some cells showed increases in PLS signals to an intermediate ridge position lying between the two ridges seen for cells prepared from oil-treated ovariectomized rats. Over the first 5 days of E2 treatment, the major effect was a progressive increase in the FALS signal of cells in this intermediate ridge position. Increases in PLS signals continued over the 12-day treatment period. Since the results of a cell sorting experiment showed that PRL cells predominated in the intermediate scatter ridge, it was concluded that E2 increases both size (FALS) and secretory granule content (PLS) of mammotrophs. Cells in the low and high scatter regions were unaffected by E2. Those in the low scatter region were shown, by both electron microscopy and light microscopic immunochemistry, to be enriched in agranular, unstained endothelial-follicular cells. The high-scatter region contained predominantly GH cells. Administration of E2 to pituitary cells cultured in serum-free or serum-containing media for 1-12 days had no effect on the FALS-PLS bivariate distribution. However, the light scatter ridges produced from cells in culture were compressed into a single ridge of narrow PLS and broad FALS signal intensity with increasing time in culture.

Flow cytometric immunofluorescence of rat anterior pituitary cells.

We have developed a flow cytometric immunofluorescence technique for the quantification of growth hormone (GH), prolactin (PRL), and luteinizing hormone (LH) producing cells. The procedure requires about 24 hours and can objectively count 50,000 cells in about 3 minutes. It is based on indirect-immunofluorescence (fluorescein) of intracellular hormone using an EPICS V cell sorter. The fluorescein distributions are gated on DNA content (propidium iodide) to eliminate counting cell clumps. Cells from the same suspensions were stained immunocytochemically and counted microscopically (1,000-2,000 cells/sample). Immunofluorescence and immunocytochemistry correlated to within a few percent for GH and PRL cells. Cell suspensions from adult males and females with or without castration and a diethylstilbestrol (DES)-induced primary pituitary tumor were used to test the method. A major finding of this study was the objective identification of two populations of PRL producing cells, i.e., lightly and intensely stained cells. On the other hand, the fluorescence distribution of PRL cells from DES-induced pituitary tumors did not fall into two distinct populations but, rather, represented a broad continuum. This method should prove useful in studying the dynamics of pituitary cell populations under various physiological and pharmacological conditions.

Electrophoretic separation of kidney and pituitary cells on STS-8.

A Continuous Flow Electrophoresis System (CFES) was used on Space Shuttle flight STS-8 to separate specific secretory cells from suspensions of cultured primary human embryonic kidney cells and rat pituitary cells. The objectives were to isolate the subfractions of kidney cells that produce the largest amounts of urokinase (plasminogen activator), and to isolate the subfractions of rat pituitary cells that secrete growth hormone, prolactin, and other hormones. Kidney cells were separated into more than 32 fractions in each of two electrophoretic runs. Electrophoretic mobility distributions in flight experiments were spread more than the ground controls. Multiple assay methods confirmed that all cultured kidney cell fractions produced some urokinase, and five to six fractions produced significantly more urokinase than the other fractions. Several fractions also produced tissue plasminogen activator. The pituitary cells were separated into 48 fractions in each of the two electrophoretic runs, and the amounts of growth hormone (GH) and prolactin (PRL) released into the medium for each cell fraction were determined. Cell fractions were grouped into eight mobility classes and immunocytochemically assayed for the presence of GH, PRL, ACTH, LH, TSH, and FSH. The patterns of hormone distribution indicate that the specialized cells producing GH and PRL are isolatable due to the differences in electrophoretic mobilities.

Pituitary hollow fiber units in vivo and in vitro.

A new technique is described for the study of mammalian pituitary cells in vivo and in vitro. The method involves encapsulation of freshly trypsinized rat, sheep or human pituitary cells in XM-50 Amicon hollow fibers followed by their intracranial implantation into hypophysectomized rats. These pituitary fiber units promoted recipient growth for 3 weeks before weight gains plateaued. Body composition analyses showed that significant quantities of protein, fat and ash accounted for the weight gain. Morphological study of the capsule contents 7-39 days postimplantation revealed the presence of intact somatotrophs and corticotrophs. The hollow fibers may provide an immunologically privileged site by virtue of the fact that the 50,000 dalton pores making up the lumen surface permit pituitary hormones to diffuse from the capsule, but theoretically do not permit immunoglobulin molecules to penetrate the capsule. Growth of hypox rats receiving capsules containing allogeneic rat pituitary cells, sheep cells or pieces of human postmortem pituitary support this concept. Furthermore, rats implanted with human PRL adenoma cells had detectable quantities of circulating hPRL 100 days postimplantation. It is suggested that the pituitary hollow fiber units function when they come in contact with a ventricular surface of a hypox animal. With these units, it will be possible to study function of the same group of pituitary cells in vitro and in vivo.

Binding of quinacrine to the human Y chromosome.

Human chromosome spreads were stained with 3H-quinacrine and their fluorescence observed. The exact location of specific spreads on each slide was noted and photographs taken. Autoradiographs were then prepared so that the quinacrine fluorescence of any specific chromosome could be compared directly with the distribution of grains over the same chromosome on the autoradiograph. The Y chromosome fluoresced much more intensely than any of the other chromosomes, but there were no more grains over the Y chromosome than over the other chromosomes. Therefore the enhanced fluorescence of the human Y chromosome is not due to an increased binding of quinacrine.