Electrolyzed Saline Targets Biofilm Periodontal Pathogens In Vitro.

Preventing the development and recurrence of periodontal diseases often includes antimicrobial mouthrinses to control the growth of the periodontal pathogens. Most antimicrobials are nonselective, targeting the symbiotic oral species as well as the dysbiosis-inducing ones. This affects the overall microbial composition and metabolic activity and consequently the host-microbe interactions, which can be detrimental (associated with inflammation) or beneficial (health-associated). Consequently, guiding the antimicrobial effect for modulating the microbial composition to a health-associated one should be considered. For such an approach, this study investigated electrolyzed saline as a novel rinse. Electrolyzed saline was prepared from sterile saline using a portable electrolysis device. Multispecies oral homeostatic and dysbiotic biofilms were grown on hydroxyapatite discs and rinsed daily with electrolyzed saline (EOS). Corresponding positive (NaOCl) and negative (phosphate-buffered saline) controls were included. After 3 rinses, biofilms were analyzed with viability quantitative polymerase chain reaction and scanning electron microscopy. Supernatants of rinsed biofilms were used for metabolic activity analysis (high-performance liquid chromatography) through measuring organic acid content. In addition, human oral keratinocytes (HOKs) were exposed to EOS to test biocompatibility (cytotoxicity and inflammation induction) and also to rinsed biofilms to assess their immunogenicity after rinsing. Rinsing the dysbiotic biofilms with EOS could reduce the counts of the pathobionts (>3 log10 Geq/mm2 reduction) and avert biofilm dysbiosis (≤1% pathobiont abundance), leading to the dominance of commensal species (≥99%), which altered both biofilm metabolism and interleukin 8 (IL-8) induction in HOKs. EOS had no harmful effects on homeostatic biofilms. The scanning electron micrographs confirmed the same. In addition, tested concentrations of EOS did not have any cytotoxic effects and did not induce IL-8 production in HOKs. EOS showed promising results for diverting dysbiosis in in vitro rinsed biofilms and controlling key periopathogens, with no toxic effects on commensal species or human cells. This novel rinsing should be considered for clinical applications.

Independent phasing of rephasing and non-rephasing 2D electronic spectra.

Assigning absolute phase to two-dimensional (2D) third-order nonlinear optical signals generally requires acquiring both the rephasing and the non-rephasing signals and comparing the sum of the two to spectrally resolved pump-probe spectra. To date, however, Gradient Assisted Photon Echo Spectroscopy (GRAPES) has only been able to acquire rephasing spectra. Such a constraint requires a new phasing protocol. Here, we analytically prove that the rephasing and non-rephasing spectra can be phased independently using pump-probe signal. We verify this result holds even for finite duration pulses by simulation. This relationship holds for all 2D spectroscopies, not only GRAPES. In addition, we present improvements to GRAPES that enable acquisition of rephasing and non-rephasing signals in different phase-matched directions. We employ our phasing protocol to phase the data for laser dye IR-144, leading to reconstruction of purely absorptive 2D spectrum.

Short communication: serum immunoglobulin G and total protein concentrations in dairy calves fed a colostrum-replacement product.

Neonatal calf health is largely dependent on the ingestion and absorption of maternally derived antibodies via colostrum administration. The objective of this study was to evaluate the efficacy of a commercially available plasma-derived colostrum-replacement (CR) product as compared with bovine colostrum. Holstein calves were removed from the dam immediately after birth and randomly allocated to 1 of 3 groups. Group 1 calves (n=22) were fed 1 package of the CR product; group 2 calves (n=22) were fed 2 packages of the CR product; and group 3 calves (n=22) were fed 3 L of bovine colostrum. Blood samples were collected from all calves 24h after colostrum or CR feeding and analyzed for serum IgG and total protein concentrations. Calves fed bovine colostrum had significantly higher serum IgG and total protein concentration than calves in either group fed the CR product. Group 1 calves (1 package of CR product) had a significantly higher incidence of failure of transfer of passive immunity than calves in groups 2 or 3. The results of this study indicated that 2 packages of this CR product achieved adequate IgG concentrations in calves. However, calves fed 1 package of this CR product consistently had failure of transfer of passive immunity.

Avian Clock gene polymorphism: evidence for a latitudinal cline in allele frequencies.

In comparison with most animal behaviours, circadian rhythms have a well-characterized molecular genetic basis. Detailed studies of circadian clock genes in 'model' organisms provide a foundation for interpreting the functional and evolutionary significance of polymorphic circadian clock genes found within free-living animal populations. Here, we describe allelic variation in a region of the avian Clock orthologue which encodes a functionally significant polyglutamine repeat (ClkpolyQcds), within free-living populations of two passerine birds, the migratory bluethroat (Luscinia svecica) and the predominantly nonmigratory blue tit (Cyanistes caeruleus). Multiple ClkpolyQcds alleles were found within populations of both species (bluethroat: 12 populations, 7 alleles; blue tit: 14 populations, 9 alleles). Some populations of both species were differentiated at the ClkpolyQcds locus as measured by F(ST) and R(ST) values. Among the blue tit, but not bluethroat populations, we found evidence of latitudinal clines in (i) mean ClkpolyQcds repeat length, and (ii) the proportions of three ClkpolyQcds genotype groupings. Parallel analyses of microsatellite allele frequencies, which are considered to reflect selectively neutral processes, indicate that interpopulation allele frequency variation at the ClkpolyQcds and microsatellite loci does not reflect the same underlying demographic processes. The possibility that the observed interpopulation ClkpolyQcds allele frequency variation is, at least in part, maintained by selection for microevolutionary adaptation to photoperiodic parameters correlated with latitude warrants further study.

The effect of image content on detail preservation and file size reduction in lossy compression.

OBJECTIVES: To demonstrate the effect of image content on image detail preservation and file size reduction. METHODS: The first set, containing 16 in vitro images with variable projection geometry, exposure time, bone level and number of teeth, was compressed with three compression modes: JPEG quality factor (JPQF), JPEG2000 quality factor (J2QF) and JPEG2000 compression ratio (J2CR). Image detail degradation was evaluated by local mean square error (MSE) on a standardized region of interest (ROI), containing bone. The second set, containing 105 clinical bitewings, was compressed with the same compression modes at 3 quality factors/compression ratios and local MSEs were calculated on two ROIs, containing bone and crown. RESULTS: For the first image set, nearly constant MSE was found for the JPQF and J2QF compression modes, while file size depended on projection geometry, exposure time, bone level and the number of teeth. In contrast, file size reduction was nearly constant for the J2CR compression mode, while MSE depended on the abovementioned factors. Similarly, for the second image set, nearly constant MSE and variation of file size reduction were found for JPQF and J2QF but not for the J2CR compression mode. All of these results were consistent for all three quality factors/compression ratios. CONCLUSIONS: Constant image detail preservation, crucial for diagnostic accuracy in radiology, can only be assured in QF compression mode in which the file size of the compressed image depends on the original image content. CR compression mode assures constant file size reduction, but image detail preservation depends on image content.

Lossy JPEG compression: easy to compress, hard to compare.

OBJECTIVES: To review the literature on lossy compression in dental radiography and to discuss the importance and suitability of the methodology used for evaluation of image compression. METHODS: A search of Medline (from 1966 to October 2004) was undertaken with the search expression "(Radiography, dental) and compression". Inclusion criterion was that the reference should be evaluating the effect of lossy image compression on diagnostic accuracy. For all included studies, information in relation to mode of image acquisition, image content, image compression, image display, and method of image evaluation was extracted. RESULTS: 12 out of 32 papers were included in the review. The design of these 12 studies was found to vary considerably. Parameters used to express the degree of information loss (DIL) were either or both compression ratio (CR) and compression level (CL). The highest acceptable CR reported in the studies ranged from 3.6% to 15.4%. Furthermore, different CR values were proposed even for the same diagnostic task, for example, for caries diagnosis CR ranged from 6.2% to 11.1%. CONCLUSION: Lossy image compression can be used in clinical radiology if it does not conflict with national law. However, the acceptable DIL is difficult to express and standardize. CR is probably not suitable to express DIL, because it is image content dependent. CL is also probably not suitable to express DIL because of the lack of compression software standardization.

DNA methylation is not likely to be responsible for aggrecan down regulation in aged or osteoarthritic cartilage.

BACKGROUND: Expression of aggrecan is reduced during aging and osteoarthritic cartilage degeneration. CpG methylation may have a role in the down regulation of aggrecan transcriptions. OBJECTIVE: To investigate whether a correlation between gene methylation and expression of aggrecan in chondrocytes exists. METHODS: The human aggrecan promoter region was analysed computationally for CpG-rich regions. These were investigated for the methylation of C residues in normal (aged) and osteoarthritic chondrocytes by the bisulphite method for modifying DNA as well as sequence analysis using DNA directly extracted from normal and osteoarthritic cartilage tissue. Additionally, chondrocytic cell lines were investigated for methylation within the aggrecan promoter region. RESULTS: The CpG-rich promoter region of the human aggrecan gene contains a 0.6 kb region that meets the criteria of a CpG island as defined by prediction programmes. A significant correlation of aggrecan mRNA expression levels and methylation status in normal (aged) and osteoarthritic chondrocytes as well as in different chondrocytic cell lines was not found. CONCLUSIONS: Expression of aggrecan in normal cartilage and diseased states is not modulated by gross changes of CpG methylation of its promoter region. CpG methylation does not have a central role in the switch off of aggrecan promoter activity in human adult articular chondrocytes.

Production of biologically active tethered ovine FSHbetaalpha by the methylotrophic yeast Pichia pastoris.

The pituitary-derived glycoprotein hormone FSH plays a central role in controlling vertebrate gonadal function. In female mammals the maturation of ovarian follicles is critically dependent upon stimulation by FSH. Moreover, injection of exogenous FSH is used extensively to stimulate increased numbers of follicles to ovulate. Structurally FSH is a heterodimeric glycoprotein composed of two non-covalently associated polypeptide subunits. The tertiary structures of both the alpha- and beta-subunits are constrained by intramolecular disulphide bonds and are post-translationally modified with two N-linked carbohydrate moieties, the structure of which appears to modulate in vivo biological activity. Here we report the expression of ovine FSH (oFSH) as a biologically active single-chain polypeptide using the methylotrophic yeast Pichia pastoris. Sequences encoding the mature oFSH alpha- and beta-proteins were fused to form a gene encoding a fusion protein with the C-terminus of the beta-chain joined to the N-terminus of the alpha-chain, with the chains separated by a two amino acid linker sequence. This fusion gene was itself fused to two alternative Pichia leader sequences (mating factor alpha and acid phosphatase) and transformed into the Pichia strains GS115 and SMD1168. The recombinant fusion protein (oFSHbetaalpha) was expressed at approximately 0.1 microg/ml in 'shake-flask' cultures. The Pichia-expressed tethered protein was biologically active in an in vitro bioassay, had a molecular mass of 28 kDa, as determined by SDS-PAGE, and bound the bovine FSH receptor with a binding profile similar to that of native oFSH.

Expression of the FcRn receptor (alpha and beta) gene homologues in the intestine of suckling brushtail possum (Trichosurus vulpecula) pouch young.

The neonatal IgG transporter FcRn consists of two chains, FcRn alpha and beta (also known as beta(2) microglobulin), and is involved in transferring IgG molecules across both mammary and intestinal epithelial cells. Developmental changes in FcRn IgG alpha and beta chain mRNA levels were investigated in the gut of brushtail possum (Trichosurus vulpecula) pouch young (PY) using Northern hybridisation. FcRn alpha transcripts were detected in the PY proximal intestine at all times examined, between days 1 and 195 of post-natal life, with increased levels detected from around day 110. The beta(2) microglobulin transcript levels in the PY proximal intestine were low to undetectable until day 110 of post-natal life and then increased dramatically after day 159. Both the FcRn alpha and beta gene transcripts were detected in a wide range of tissues in the adult possum (>365 days). Genomic sequences located 5' to the start of transcription of the FcRn alpha and beta(2) microglobulin genes were cloned and analysed for predicted cis-acting transcription control elements. Both the FcRn alpha and beta(2) microglobulin genomic sequences contained STAT5 binding motifs consistent with the transcription of both genes being modulated by prolactin. Using in situ hybridisation, the FcRn alpha and beta(2) microglobulin transcripts were localised to the epithelial cells of the PY intestine. However, no prolactin receptor transcripts were detected in the same epithelial cells suggesting that the observed changes in FcRn alpha and beta(2) microglobulin gene expression in the proximal intestine are not modulated directly by prolactin. The results are consistent with the hypothesis that changes in FcRn alpha and beta(2) microglobulin gene expression take place in the possum PY intestine to accommodate changes in maternal milk composition to meet the changing immunological demands of the PY.

Comparative evaluation of JPEG and JPEG2000 compression in quantitative digital subtraction radiography.

OBJECTIVES: The aim of this in vitro study was to compare the impact of JPEG and the novel JPEG2000 compression standard on quantitative digital subtraction radiography (DSR) and to determine the acceptable JPEG2000 compression ratios for DSR. METHODS: Nine dry pig mandible sections were radiographed three times ('Baseline', 'No change', and 'Gain') with standardized projection geometry. Bone gain was simulated by adding artificial bone chips (1, 4 and 15 mg). Images were registered, compressed by JPEG and JPEG2000 using compression ratios (CR) of 1 : 7, 1 : 16, 1 : 22, and 1 : 31, and then subtracted. Image distortion was assessed objectively by calculating average pixel error and peak signal to noise ratio. No change areas in compressed and subtracted 'No change-Baseline' images and bone gain volumes in compressed and subtracted 'Gain-Baseline' images were calculated for both compression standards and compared. RESULTS: JPEG introduced less distortion at low CRs, while JPEG2000 was superior at higher CRs. At CR of 1 : 7, no significant difference between JPEG and JPEG2000 was found. JPEG2000 yielded better results for no change measurements at higher CRs. Volumes of simulated bone gain were overestimated when JPEG and underestimated when JPEG2000 compression was used. CONCLUSIONS: At CR of 1 : 7 JPEG and JPEG2000 performed similarly, which indicates that CR of 1:7 in JPEG2000 can be used for DSR if images are registered before compression. At higher CRs, JPEG2000 is superior to JPEG but image distortions are too high for reliable quantitative DSR.

Impact of JPEG lossy image compression on quantitative digital subtraction radiography.

OBJECTIVES: The aim of the study was to evaluate the impact of JPEG lossy image compression on the estimation of alveolar bone gain by quantitative digital subtraction radiography (DSR). METHODS: Nine dry domestic pig mandible posterior segments were radiographed three times ('Baseline', 'No change', and 'Gain') with standardized projection geometry. Bone gain was simulated by adding artificial bone chips (1, 4, and 15 mg). Images were either compressed before or after registration. No change areas in compressed and subtracted 'No change-Baseline' images and bone gain volumes in compressed and subtracted 'Gain-Baseline' images were calculated and compared to the corresponding measurements performed on original subtracted images. RESULTS: Measurements of no change areas ('No change-Baseline') were only slightly affected by compressions down to JPEG 50 (J50) applied either before or after registration. Simulated gain of alveolar bone ('Gain-Baseline') was underestimated when compression before registration was performed. The underestimation was bigger when small bone chips of 1 mg were measured and when higher compression rates were used. Bone chips of 4 and 15 mg were only slightly underestimated when using J90, J70, and J50 compressions before registration. CONCLUSIONS: Lossy JPEG compression does not affect the measurements of no change areas by DSR. Images undergoing subtraction should be registered before compression and if so, J90 compression with a compression ratio of 1:7 can be used to detect and measure 4 mg and larger bone gain.

Residual neurologic deficits 30 years after occupational exposure to elemental mercury.

A battery of tests of peripheral and central nervous system function was administered to 205 former workers of a large heavy industrial plant, 104 of whom were previously exposed to inorganic mercury. The mean age of those examined was 71 years. Exposed subjects had participated in a urine-mercury exposure monitoring program during the time of operation of a process that required the use of mercury and its subsequent clean-up. Mercury exposure had been high (mean peak urine mercury concentration was >600 microg/l) and had ended 30 years or more prior to the investigation. Peripheral nerve function outcomes that were statistically significantly associated with cumulative mercury exposure after controlling for covariates included classification as having peripheral neuropathy, peroneal motor nerve conduction velocity, ulnar motor nerve conduction velocity, and peroneal motor nerve F-wave latency. Quantitative assessment of resting tremor was nearly significantly associated with cumulative mercury exposure (p=0.07). Among tests of central nervous system function, results of the Handeye Coordination test were significantly associated with cumulative mercury exposure after controlling for covariates. Cumulative mercury exposure was not observed to be associated with a quantitative measure of dementia or with a number of cognitive neurobehavioral test outcomes. The statistically significant associations with mercury exposure were observed in spite of greater mortality among the exposed group than the unexposed group. These results suggest that substantial occupational mercury exposure can have long-term adverse effects on the peripheral nervous system detectable decades after cessation of exposure. Such long-term adverse effects were not observed for a measure of dementia or other measures of cognitive function.

Growth and paracrine factors regulating follicular formation and cellular function.

The purpose of this paper is to review, using fetal sheep as the animal model, aspects of ovarian development related to follicular formation and to report on the identity of growth and paracrine factors which might be involved in this process. Before follicular formation there is a massive and sustained colonisation of the fetal ovary by mesonephric cells, which become a precursor source of follicular cells. From within the ovarian medulla, somatic 'cell-streams' branch into the cortex around nests of oogonia and oocytes. These 'cell-streams', which contain elongated cells with either flattened or cuboidal shaped nuclei, express steroidogenic factor-1 (SF-1), steroid acute regulatory protein (StAR), 3beta-hydroxysteroid dehydrogenase (3beta-HSD), cytochrome P450(scc), and P450(aromatase) mRNA and/or protein. Follicles form from the association of an oocyte with the 'cell-stream' with either a single layer of flattened cells (i.e. type 1 follicle) or with a mixture of flattened and cuboidal cells (i.e. type 1a follicle). These newly-formed follicles have between 3 and 57 somatic cells (i.e. granulosa cells) and contain oocytes which vary in diameter between 23 and 52 microm. Newly formed and early growing follicles have been identified with growth factors or growth factor receptors in either the oocytes or granulosa cells. Many of the growth factors are from the TGFbeta superfamily and are expressed in a cell- and stage-specific manner.

Influence of developer exhaustion on accuracy of quantitative digital subtraction radiography: an in vitro study.

OBJECTIVE: The aim of the study was to evaluate the influence of developer exhaustion on accuracy of quantitative digital subtraction radiography. STUDY DESIGN: Six objects, each incorporating a section of dry human mandible, were radiographed with 4 exposure times. Baseline films were processed in fresh solutions, whereas follow-up films were processed in fresh and in increasingly exhausted solutions (ie, 1, 2, and 3 weeks old). Bone loss and bone gain were computer simulated in 17 regions of interest on baseline radiographs. Area and volume of changes in mineralization were measured in subtracted images, obtained by subtraction of baseline from their corresponding follow-up radiographs. Friedman's 2-way analysis of variance by ranks and Wilcoxon signed-rank test were used for statistical analysis. RESULTS: Because of exhausted developer, bone loss was relatively underestimated from 6.6% to 16.5% (P <.05), whereas bone gain was relatively overestimated from 9.7% to 16.7% (P <.05). CONCLUSIONS: This in vitro study demonstrates that films for quantitative digital subtraction radiography should be processed in fresh developer or error might be introduced.

Cloning and expression, pharmacological characterization, and internalization kinetics of the pituitary GnRH receptor in a metatherian species of mammal.

Gonadotropin-releasing hormone receptors (GnRH-Rs) expressed in the pituitary of eutherian species of mammal are unique in lacking the cytoplasmic C-terminal tail characteristic of GnRH-Rs of nonmammalian vertebrates and other G protein-coupled receptors. To further investigate evolutionary relationships among vertebrate GnRH-Rs, a full-coding region cDNA of the pituitary GnRH-R was cloned from a metatherian marsupial mammal, the Australian brushtail possum (Trichosurus vulpecula). We have determined the pharmacological characteristics and internalization kinetics of this GnRH-R from an early evolved, metatherian species of mammal and compared it with the corresponding receptors in eutherian species of mammal and nonmammalian vertebrates. The predicted GnRH-R protein from the possum pituitary has high homology with the other mammalian GnRH-Rs (80% identity) and, in common with other mammals, lacks an intracellular C-terminal tail. The ligand selectivity of the possum GnRH-R transfected into COS-1 cells, assessed using inositol phosphate assays and radioreceptor binding assays, was similar to that of the other mammalian GnRH-Rs, and distinct from those of the nonmammalian GnRH-Rs. The pharmacological characteristics of the possum GnRH-R were similar to those of other mammalian GnRH-Rs, for a selection of agonists (including naturally occurring GnRH ligands and superagonists) and antagonists. Receptor-mediated internalization of GnRH agonist by the possum GnRH-R was slightly more rapid than that of the human GnRH-R, while the internalization kinetics of the chicken GnRH-R, in which a cytoplasmic C-terminal tail is present, was considerably more rapid. In terms of the evolution of the GnRH-R in vertebrates, the possum (a metatherian mammal) GnRH-R has a striking resemblance, in both structure and pharmacological characteristics, to GnRH-Rs in eutherian mammals, which are quite distinct from the nonmammalian vertebrate GnRH-Rs, and are unique among G protein-coupled receptors in lacking an intracellular C-terminal tail. The distinct structure of the pituitary GnRH-R in mammalian vertebrates is likely to have important functional consequences in the reproductive physiology of mammals.

Screening the foods of an endangered parrot, the kakapo (Strigops habroptilus), for oestrogenic activity using a recombinant yeast bioassay.

In recent years the possibility of environmental oestrogens affecting the reproduction of vertebrates has become an issue of both public and scientific interest. Although the significance of such chemicals remains controversial there is clear evidence that, in some contexts, environmental oestrogens can influence the fertility of vertebrates. Highly endangered species represent a situation in which even modest reductions in the fertility of key individuals may have implications for the survival of the entire species. This paper reports the screening of both natural and supplementary foods of the kakapo (Strigops habroptilus), a critically endangered New Zealand nocturnal parrot, for oestrogenic activity using a recombinant yeast based bioassay. Low levels of oestrogenic activity were detected in one of the 'chick-raising' foods, but no oestrogenic activity was detected in the adult supplementary foods. The oestrogenicity of a range of phytochemicals possibly associated with the kakapo natural diet was also examined. Two such phytochemicals, podocarpic acid and its reduced derivative podocarpinol, showed weak oestrogenic activity (approximately 10(-6) and 10(-4) of the activity of 17-beta-oestradiol, respectively).

Stem cell factor and c-kit gene expression and protein localization in the sheep ovary during fetal development.

The aim of this study was to investigate stem cell factor and c-kit gene expression and protein localization in the mesonephros and ovary of sheep fetuses at different days of gestation, using RNA in situ hybridization and immunohistochemical procedures. At days 24 and 26 of gestation, stem cell factor mRNA and protein were present in cells throughout the developing gonad and mesonephros. From day 28 to day 40 of gestation, stem cell factor mRNA and protein became increasingly localized to the cortical region of the ovary, where most germ cells were present as actively proliferating oogonia. From day 40 to day 90 of gestation, stem cell factor mRNA and protein localization were confined mainly to the ovarian cortex. Moreover, within the cortical region, stem cell factor mRNA was low or absent where follicles were first forming and highest in the outer ovarian cortex, where germ cells were undergoing mitosis or the early stages of meiosis. In contrast, stem cell factor protein was present in newly forming follicles, as well as in mitotic and meiotic germ cells, which is consistent with the presence of both membrane-bound and soluble forms of this ligand. However, by day 90 of gestation, both stem cell factor mRNA and protein were observed in the granulosa cells of most (> 90%) primordial follicles. C-kit mRNA and protein were observed from day 24 of gestation in both germ cells and somatic cells but, with increasing gestational age, preferentially in germ cells (for example, pre-meiotic germ cells and both isolated oocytes and follicle-enclosed oocytes). C-kit protein, but not mRNA, was also observed in germ cells that were undergoing meiosis. The results indicate that the cells containing stem cell factor mRNA within the ovary up to day 90 of gestation originated from the gonadal blastema and from cells that migrated from the mesonephros before day 28 of gestation. Since stem cell factor mRNA was absent in both mesonephric cells migrating after day 28 of gestation and in regions where follicles were first forming, it is suggested that these later migrating mesonephric cells are the progenitors of the granulosa cells in the first forming follicles. In conclusion, during follicle formation, c-kit mRNA is localized to germ cells whereas c-kit, together with stem cell factor protein, is localized to both germ cells and somatic cells, consistent with the hypothesis that the presence of this receptor-ligand pair is essential to prevent apoptosis.

Control of early ovarian follicular development.

Early follicular growth refers to the development of an ovarian follicle from the primordial to early antral phase. In sheep and cows these phases of growth can be classified by the configuration of granulosal cells in the largest cross-section of the follicle as types 1 (primordial), 1a (transitory) 2 (primary), 3 and 4 (preantral) and 5 (early antral). Follicles classified as type 1 may be highly variable within each species with respect to number of granulosal cells and diameter of oocyte. Much of the variation in granulosal cell composition of type 1 follicles may occur at formation and this may account for the variability in granulosal cell composition throughout subsequent stages of growth. There appear to be important differences among species (for example sheep and cattle) in the number and function of granulosal cells relative to the diameter of the oocyte during the initiation of follicular growth. There is evidence that most, if not all, of the growth phases from types 1 to 5 are gonadotrophin-independent and that follicles develop in a hierarchical manner. In sheep, cows and pigs, numerous growth factor, growth factor receptor and gonadotrophin receptor mRNAs and peptides (for example c-kit, stem cell factor, GDF-9, beta B and beta A activin/inhibin subunit, alpha inhibin subunit, follistatin, FGF-2, EGF, EGF-R, TGF beta 1,2 and 3 FSH-R and LH-R) are expressed in a phase of growth (for example types 1-5)-specific and cell-specific manner. However, the roles of many of these factors remain to be determined.

Infertility: from a personal to a public health problem.

The inability to conceive a child is most often viewed as a private matter, but public health perspectives and skills can contribute greatly to our knowledge about infertility, and the development of effective and rational public policy for prevention, access to health care, and regulation of new technologies. We offer a primer of public health aspects of infertility in an effort to encourage the broad spectrum of public health professionals to become more knowledgeable about these topics and join in the national debate about preventive strategies, cost-benefit assessment, resource allocation, and ethics.