A 31P topical magnetic resonance study of embryonic development in hens' eggs

1. A ³¹P topical magnetic resonance study of whole chicken's eggs in vivo is described.

2. Resonances from adenosine triphosphate, phosphocreatine, inorganic phosphate and phosphorous storage protein were observed. These changed in relative intensity as the embryo developed.

3. No ill effects due to exposure to radiofrequency radiation or magnetic fields were observed.

4. It is concluded that topical magnetic resonance can make a useful contribution to the study of embryonic development in avian eggs.     

TDP-43 mutations in familial and sporadic amyotrophic lateral sclerosis

Amyotrophic lateral sclerosis (ALS) is a fatal motor neuron disorder characterized pathologically by ubiquitinated TAR DNA binding protein (TDP-43) inclusions. The function of TDP-43 in the nervous system is uncertain, and a mechanistic role in neurodegeneration remains speculative. We identified neighboring mutations in a highly conserved region of TARDBP in sporadic and familial ALS cases. TARDBPM337V segregated with disease within one kindred and a genome-wide scan confirmed that linkage was restricted to chromosome 1p36, which contains the TARDBP locus. Mutant forms of TDP-43 fragmented in vitro more readily than wild type and, in vivo, caused neural apoptosis and developmental delay in the chick embryo. Our evidence suggests a pathophysiological link between TDP-43 and ALS.     

Probing the Local Effects of Magnetic Impurities on Superconductivity

The local effects of isolated magnetic adatoms on the electronic properties of the surface of a superconductor were studied with a low-temperature scanning tunneling microscope. Tunneling spectra obtained near magnetic adsorbates reveal the presence of excitations within the superconductor's energy gap that can be detected over a few atomic diameters around the impurity at the surface. These excitations are locally asymmetric with respect to tunneling of electrons and holes. A model calculation based on the Bogoliubov-de Gennes equations can be used to understand the details of the local tunneling spectra.     

Idiopathic hematuria of unilateral renal origin in a dog

Idiopathic hematuria of unilateral renal origin was diagnosed in a 3-month-old male Weimaraner. The hematuria had been visible grossly for 6 weeks. Extensive clinicopathologic, radiographic, and histologic evaluations localized the source of the hematuria to the right kidney, but could not identify a cause. Nephrectomy of the right kidney resolved the problem. This case report supports a previous article comparing canine idiopathic hematuria of renal origin with benign essential hematuria of human beings.     

Recent pine woodland dynamics in east Glen Affric, northern Scotland, from highly resolved palaeoecological analyses

Glen Affric, a National Nature Reserve of international conservationimportance for plant and animal communities associated withScots pine (Pinus sylvestris) woodland is managed for natureconservation and woodland restoration at a landscape scale.Management plans have drawn on information on current standstructure and variation but have not used in any detailed waythe analyses of past tree population changes from sources, suchas pollen analyses. This paper reports the results of pollenanalyses from three small peat hollows at the head of Loch Beinna' Mheadhoin. The analyses demonstrate, first, that woods inthe east of the reserve several centuries ago were differentin species composition and were more varied than they are todayand, second, that the currently patchy Pinus wood in the westof the reserve, at the head of Loch Beinn a' Mheadhoin, is theresult of recent spread westward onto former heathland. Thistemporal and spatial variability in the recent past has implicationsfor the future management of the woods because future woodsmay not develop with the characteristics of the current stands,and may not be stable over time.     

Altering the substrate specificity of Candida rugosa LIP4 by engineering the substrate-binding sites

Candida rugosa (formerly Candida cylindracea) lipase (CRL) is an important industrial enzyme that is widely used in biotechnological applications such as the production of fatty acids and the synthesis of various esters. CRL comprises at least seven isozymes (LIP1-LIP7), which share a similar amino acid sequence but with different specificities for substrates. Previously, LIP4 was reported to have higher esterase activity toward long acyl-chain ester and lower lipase activity toward triglycerides. A296 and V344 of LIP4 were predicted to play decisive roles in its substrate specificity. In this study, site-specific saturation mutagenesis has been employed to study the substrate specificity of LIP4. Point mutations were separately introduced into A296 and V344 positions using degenerate primer sets containing 32 codons to generate two libraries of variants. LIP4 variants were heterologously expressed in the yeast Pichia pastoris. A specific plate assay was used to identify lipase-producing P. pastoris clones in a medium containing tributyrin. LIP4 variants with high activity toward short fatty acyl-chain triglyceride (tributyrin) were screened. Specificity analysis and biochemical characterization indicated that the recombinant variants A296I, V344Q, and V344H had properties remarkably different from those of wild-type LIP4. All three variant enzymes had significantly higher specific activities toward tributyrin than LIP4. In addition to short-chain triglyceride, A296I and V344Q also improved hydrolytic activities of triglycerides toward medium- and long-chain triglycerides tested. The results suggested that A296 played an important role in lipase activity and high-temperature dependence of LIP4, whereas it had no effect on the chain-length specificity in lipolytic reaction. The V344 residue had a significant effect on the substrate chain-length specificity of LIP4.     

Use of Fourier‐Transform Infrared Spectroscopy to Quantify Immunoglobulin G Concentrations in Alpaca Serum

Background

Rapid, economical, and quantitative assays for measurement of camelid serum immunoglobulin G (IgG) are limited. In camelids, failure of transfer of maternal immunoglobulins has a reported prevalence of up to 20.5%. An accurate method for quantifying serum IgG concentrations is required.

Objective

To develop an infrared spectroscopy‐based assay for measurement of alpaca serum IgG and compare its performance to the reference standard radial immunodiffusion (RID) assay.

Animals

One hundred and seventy‐five privately owned, healthy alpacas.

Methods

Eighty‐two serum samples were collected as convenience samples during routine herd visits whereas 93 samples were recruited from a separate study. Serum IgG concentrations were determined by RID assays and midinfrared spectra were collected for each sample. Fifty samples were set aside as the test set and the remaining 125 training samples were employed to build a calibration model using partial least squares (PLS) regression with Monte Carlo cross validation to determine the optimum number of PLS factors. The predictive performance of the calibration model was evaluated by the test set.

Results

Correlation coefficients for the IR‐based assay were 0.93 and 0.87, respectively, for the entire data set and test set. Sensitivity in the diagnosis of failure of transfer of passive immunity (FTPI) ([IgG] <1,000 mg/dL) was 71.4% and specificity was 100% for the IR‐based method (test set) as gauged relative to the RID reference method assay.

Conclusions and Clinical Importance

This study indicated that infrared spectroscopy, in combination with chemometrics, is an effective method for measurement of IgG in alpaca serum.