(1) Occult and latent tuberculous infection of mesenteric lymph glands; and, (2) Description of two stretcher splints for the non-operative treatment of bone or joint tuberculosis with notes on cases treated, photographs and sketches

(1) Occult and latent tuberculous infection of mesenteric lymph glands:On histological evidence alone then, this series goes to prove that, as in the upper digestive tract so also in the lower, tubercle bacilli may pass through intact mucous membrane without leaving a trace of their pathway, be arrested in the nearest lymph glands, and there lie latent and occult for a long period awaiting the opportunity for further progression.(2) Description of two stretcher splints for the non-operative treatment of bone or joint tuberculosis with notes on cases treated, photographs and sketches:SPINAL TUBERCULOSIS • HIP-JOINT TUBERCULOSIS • KNEE -JOINT TUBERCULOSIS • THE PROBLEM OF DROP FOOT • A PORTABLE SPLINT FOR THE CORRECTION OF LATERAL DEVIATION OR SCOLIOSIS OF THE SPINE

DIFFERENTIAL EFFECTS OF IGF-I AND IGF-II ON THE FATE OF MOUSE TROPHOBLAST STEM CELLS

The main objective of this thesis was to identify the role(s) of IGF-I and IGF-II on the fate of mouse trophoblast stem cells (TS cells) and their role in contributing to the stem cell fate, when grown in the absence of additional growth factors. By analyzing the expression of self-renewal and trophoblast-specific transcription factors, the addition of IGF-I and IGF-II restored nuclear OCT4 and CDX2 expression, with IGF- II inducing a longer effect, while the expression of SOX2 was not restored. IGF-I was found to induce the differentiation of TS cells based on expression and quantitative analysis of PL-1 expression. Annexin-V and TUNEL staining demonstrated that IGF-II protects TS cells from cell death. The addition of FGF-4 to the system did not have an additive effect of protection and induced further negative effect when combined with IGFs. These studies demonstrate that IGF-I and IGF-II, have different biological effects on the fate of mouse TS cells, likely through different signal transduction pathways

Evolution in the genus Arum: a comparative analysis of morpohological and genetic variation.

Testing the correlation of morphological and genetic marker variation enables the investigation of evolutionary processes. Knowledge of evolutionary processes can be used to identify those morphological characters that could be used to produce evolutionary meaningful taxonomies. This thesis aims to test the correlation between morphological and genetic marker variation to further understand the evolution of species within the genus Arum and identify those morphological characters that correspond with evolutionary groups. The investigation is carried out at the intraspecific level, intrageneric level and in a putative hybrid zone. At the intraspecific level, genetic (ISSR) and morphological variation was quantified in populations of the morphologically similar species A. maculatum and A. italicum. Populations of A. maculatum showed evidence of isolation by distance, presumably a result of pollinator behaviour and seed dispersal. Leaf patterning in A. maculatum did not correspond to evolutionary lineages. However, similar leaf patterning characters in A. italicum are used to classify the two subspecies neglectum and italicum and the ISSR analysis confirmed that these taxa are genetically distinct. These two subspecies were shown to be interbreeding in sympatric populations. The interbreeding has created a morphological and genetic difference between subsp. neglectum in sympatric populations compared with allopatric populations. At the intrageneric level, a phylogenetic analysis of Arum (using trnL and ITS 1 sequences) indicated that both vegetative and reproductive characters are convergent within the genus. The apparent convergent evolution of reproductive and vegetative characters indicates that both have been important during the diversification of the genus. These convergent characters are not useful for producing classifications that reflect evolutionary groups as the groups they produce are polyphyletic. In the putative hybrid zone, ISSR markers confirmed the presence of A. creticum and A. idaeum hybrids. There appears to be introgression of the A. idaeum genome into A. creticum; this could have implications for the future genetic integrity of A. creticum. Within this hybrid zone, continuous characters were found to be representative of genetic variation, however categorical characters were not. In conclusion, this thesis has shown that even within a single genus, the correlation between morphological and genetic marker variation is influenced by both the taxa being studied and the nature of the morphological trait. In particular, if morphological characters are found to be adaptively important, their correspondence to genetic groups should be tested before their use in taxonomies. The findings of this thesis also suggest there is great value in the complementary use of genetic and morphological analysis for taxonomic studies as well as evolutionary studies. For example, the importance of reproductive characters in the diversification of Arum species has produced a wide range of morphological variation, with limited taxonomic utility due to a tendency for homoplasy. Vegetative characters were also found to need careful testing before use in taxonomies as leaf patterning was found to correspond to sub-species status for one species of Arum but not another. Finally, this thesis has shown that, if closely related taxa are hybridising, variation of continuous reproductive characters may be used as an indicator of hybridisation, even if the morphological characters are potentially polygenic

Evolution in the genus Arum : a comparative analysis of morphological and genetic variation

Testing the correlation of morphological and genetic marker variation enables the investigation of evolutionary processes. Knowledge of evolutionary processes can be used to identify those morphological characters that could be used to produce evolutionary meaningful taxonomies. This thesis aims to test the correlation between morphological and genetic marker variation to further understand the evolution of species within the genus Arum and identify those morphological characters that correspond with evolutionary groups. The investigation is carried out at the intraspecific level, intrageneric level and in a putative hybrid zone. At the intraspecific level, genetic (ISSR) and morphological variation was quantified in populations of the morphologically similar species A. maculatum and A. italicum. Populations of A. maculatum showed evidence of isolation by distance, presumably a result of pollinator behaviour and seed dispersal. Leaf patterning in A. maculatum did not correspond to evolutionary lineages. However, similar leaf patterning characters in A. italicum are used to classify the two subspecies neglectum and italicum and the ISSR analysis confirmed that these taxa are genetically distinct. These two subspecies were shown to be interbreeding in sympatric populations. The interbreeding has created a morphological and genetic difference between subsp. neglectum in sympatric populations compared with allopatric populations. At the intrageneric level, a phylogenetic analysis of Arum (using trnL and ITS 1 sequences) indicated that both vegetative and reproductive characters are convergent within the genus. The apparent convergent evolution of reproductive and vegetative characters indicates that both have been important during the diversification of the genus. These convergent characters are not useful for producing classifications that reflect evolutionary groups as the groups they produce are polyphyletic. In the putative hybrid zone, ISSR markers confirmed the presence of A. creticum and A. idaeum hybrids. There appears to be introgression of the A. idaeum genome into A. creticum; this could have implications for the future genetic integrity of A. creticum. Within this hybrid zone, continuous characters were found to be representative of genetic variation, however categorical characters were not. In conclusion, this thesis has shown that even within a single genus, the correlation between morphological and genetic marker variation is influenced by both the taxa being studied and the nature of the morphological trait. In particular, if morphological characters are found to be adaptively important, their correspondence to genetic groups should be tested before their use in taxonomies. The findings of this thesis also suggest there is great value in the complementary use of genetic and morphological analysis for taxonomic studies as well as evolutionary studies. For example, the importance of reproductive characters in the diversification of Arum species has produced a wide range of morphological variation, with limited taxonomic utility due to a tendency for homoplasy. Vegetative characters were also found to need careful testing before use in taxonomies as leaf patterning was found to correspond to sub-species status for one species of Arum but not another. Finally, this thesis has shown that, if closely related taxa are hybridising, variation of continuous reproductive characters may be used as an indicator of hybridisation, even if the morphological characters are potentially polygenic.EThOS - Electronic Theses Online ServiceGBUnited Kingdo

Highly redundant neuropeptide volume co-transmission underlying episodic activation of the GnRH neuron dendron.

The necessity and functional significance of neurotransmitter co-transmission remains unclear. The glutamatergic 'KNDy' neurons co-express kisspeptin, neurokinin B (NKB), and dynorphin and exhibit a highly stereotyped synchronized behavior that reads out to the gonadotropin-releasing hormone (GnRH) neuron dendrons to drive episodic hormone secretion. Using expansion microscopy, we show that KNDy neurons make abundant close, non-synaptic appositions with the GnRH neuron dendron. Electrophysiology and confocal GCaMP6 imaging demonstrated that, despite all three neuropeptides being released from KNDy terminals, only kisspeptin was able to activate the GnRH neuron dendron. Mice with a selective deletion of kisspeptin from KNDy neurons failed to exhibit pulsatile hormone secretion but maintained synchronized episodic KNDy neuron behavior that is thought to depend on recurrent NKB and dynorphin transmission. This indicates that KNDy neurons drive episodic hormone secretion through highly redundant neuropeptide co-transmission orchestrated by differential post-synaptic neuropeptide receptor expression at the GnRH neuron dendron and KNDy neuron

Highly redundant neuropeptide volume co-transmission underlying episodic activation of the GnRH neuron dendron.

The necessity and functional significance of neurotransmitter co-transmission remains unclear. The glutamatergic 'KNDy' neurons co-express kisspeptin, neurokinin B (NKB), and dynorphin and exhibit a highly stereotyped synchronized behavior that reads out to the gonadotropin-releasing hormone (GnRH) neuron dendrons to drive episodic hormone secretion. Using expansion microscopy, we show that KNDy neurons make abundant close, non-synaptic appositions with the GnRH neuron dendron. Electrophysiology and confocal GCaMP6 imaging demonstrated that, despite all three neuropeptides being released from KNDy terminals, only kisspeptin was able to activate the GnRH neuron dendron. Mice with a selective deletion of kisspeptin from KNDy neurons failed to exhibit pulsatile hormone secretion but maintained synchronized episodic KNDy neuron behavior that is thought to depend on recurrent NKB and dynorphin transmission. This indicates that KNDy neurons drive episodic hormone secretion through highly redundant neuropeptide co-transmission orchestrated by differential post-synaptic neuropeptide receptor expression at the GnRH neuron dendron and KNDy neuron

Identification of influential probe types in epigenetic predictions of human traits: implications for microarray design

BACKGROUND: CpG methylation levels can help to explain inter-individual differences in phenotypic traits. Few studies have explored whether identifying probe subsets based on their biological and statistical properties can maximise predictions whilst minimising array content. Variance component analyses and penalised regression (epigenetic predictors) were used to test the influence of (i) the number of probes considered, (ii) mean probe variability and (iii) methylation QTL status on the variance captured in eighteen traits by blood DNA methylation. Training and test samples comprised ≤ 4450 and ≤ 2578 unrelated individuals from Generation Scotland, respectively. RESULTS: As the number of probes under consideration decreased, so too did the estimates from variance components and prediction analyses. Methylation QTL status and mean probe variability did not influence variance components. However, relative effect sizes were 15% larger for epigenetic predictors based on probes with known or reported methylation QTLs compared to probes without reported methylation QTLs. Relative effect sizes were 45% larger for predictors based on probes with mean Beta-values between 10 and 90% compared to those based on hypo- or hypermethylated probes (Beta-value ≤ 10% or ≥ 90%). CONCLUSIONS: Arrays with fewer probes could reduce costs, leading to increased sample sizes for analyses. Our results show that reducing array content can restrict prediction metrics and careful attention must be given to the biological and distribution properties of CpG probes in array content selection. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13148-022-01320-9

Definition of Estrogen Receptor Pathway Critical for Estrogen Positive Feedback to Gonadotropin-Releasing Hormone Neurons and Fertility

SummaryThe mechanisms through which estrogen regulates gonadotropin-releasing hormone (GnRH) neurons to control mammalian ovulation are unknown. We found that estrogen positive feedback to generate the preovulatory gonadotropin surge was normal in estrogen receptor β knockout (ERβ) mutant mice, but absent in ERα mutant mice. An ERα-selective compound was sufficient to generate positive feedback in wild-type mice. As GnRH neurons do not express ERα, estrogen positive feedback upon GnRH neurons must be indirect in nature. To establish the cell type responsible, we generated a neuron-specific ERα mutant mouse line. These mice failed to exhibit estrogen positive feedback, demonstrating that neurons expressing ERα are critical. We then used a GnRH neuron-specific Pseudorabies virus (PRV) tracing approach to show that the ERα-expressing neurons innervating GnRH neurons are located within rostral periventricular regions of the hypothalamus. These studies demonstrate that ovulation is driven by estrogen actions upon ERα-expressing neuronal afferents to GnRH neurons