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Biomedical subjects

K M Green

Publications and source records attributed to K M Green.

At least 19 recordsLinked to original sources

Proteomic analysis of human meibomian gland secretions.

BACKGROUND/AIM: Human tears contain hundreds of proteins that may exert a significant influence on tear film stability, ocular surface integrity, and visual function. The authors hypothesise that many of these proteins originate from the meibomian gland. This study's aim was to begin to develop the proteomic methodology to permit the testing of their hypothesis. METHODS: Meibomian gland secretions were collected from the lower eyelids of adult volunteers and placed in a chloroform-methanol mixture. Samples were partitioned in a biphasic system and non-lipid phase materials were reduced, alkylated, and trypsin digested to obtain peptides for protein identification. This peptide mixture was separated by micro-capillary reverse phase chromatography and the effluent examined by nano-electrospray MS and data dependent MS/MS. SEQUEST software was used to identify proteins from the MS/MS spectra. RESULTS: The methodological approach to date has permitted the identification of more than 90 proteins in human meibomian gland secretions. Proteins include the alpha2-macroglobulin receptor, IgA alpha chain, farnesoid X activated receptor, interferon regulatory factor 3, lacritin precursor, lactotransferrin, lipocalin 1, lysozyme C precursor, potential phospholipid transporting ATPase IK, seven transmembrane helix receptor (also termed somatostatin receptor type 4), testes development related NYD-SP21 (also termed high affinity IgE receptor beta subunit), and TrkC tyrosine kinase. CONCLUSIONS: These findings indicate that the meibomian gland secretes a number of proteins into the tear film. It is quite possible that these proteins contribute to the dynamics of the tear film in both health and disease.

Adult↗

Reduction of nasal fractures under local anaesthetic.

Forty five patients with fractured nasal bones underwent nasal fracture reduction under local anaesthetic. Patients used linear analog scores to assess pain associated with the procedure and satisfaction with the reduction. The surgeon also recorded his assessment of the reduction. The patients were reviewed two weeks later and the outcome scores repeated. The procedure was well tolerated and forty three patients (95.5%) stated they would undergo the same procedure again if they fractured their nose in the future. Fifteen patients (33.3%) had significant persistent nasal deformities and were listed for septorhinoplasty. The results of nasal fracture reduction under localanaesthetic are comparable to those obtained under general anaesthetic. The factors that may contribute to poor outcomes following reduction of nasal fractures are discussed.

Adolescent↗

Amyloidosis of Waldeyer's ring and larynx.

Amyloidosis of the upper aerodigestive tract is relatively rare. A case of localized amyloidosis involving all components of Waldeyer's ring with added laryngeal involvement is described. This has not been previously reported. A literature review of this conditions is presented.

Amyloidosis↗

Are pre-operative vocal fold checks necessary?

Over a 26-year period, from 1970 to 1995, 2250 pre-operative vocal fold checks were carried out by the ENT team at Manchester Royal Infirmary in patients undergoing thyroid, parathyroid and cardiothoracic surgery. A register was kept of all pre-operative fold checks. This register was retrospectively analysed and 23 vocal fold palsies were documented. In six of these patients the notes were either missing or incomplete. All of the remaining 17 patients had a history suggestive of vocal fold palsy. We suggest that in the absence of symptoms, either current or on history, a routine pre-operative vocal fold check is unnecessary.

Adult↗

Alar haematoma.

We present two cases of alar haematoma. This is a very rare complication of nasal trauma, and only two cases have been described previously. One case presented late and did not undergo surgical drainage and has a persistent cosmetic deformity. We recommend early surgical drainage when possible.

Adult↗

Non-cardiogenic pulmonary oedema due to foreign body aspiration.

A patient who developed non-cardiogenic pulmonary oedema secondary to acute airway obstruction caused by an aspirated foreign body is presented. The literature is reviewed, discussing the theories regarding the formation of non-cardiogenic pulmonary oedema. The case highlights the importance of this rare complication of foreign body aspiration and surgeons and anaesthetists should be alert to continued respiratory symptoms following relief of acute airway obstruction.

Airway Obstruction↗

Effect of myosin light chain kinase, protein kinase A, and protein kinase C inhibition on porcine oocyte activation.

Previous studies have shown that exposure to broad-spectrum protein kinase inhibitors results in parthenogenetic activation of metaphase II arrested porcine oocytes. The objective of this study was to determine the effect of inhibitors of myosin light chain kinase and other protein kinases on pronuclear development, dephosphorylation of a 25-kDa protein, and cortical granule exocytosis. Metaphase II arrested oocytes were obtained by in vitro maturation. Cumulus-free oocytes were cultured with specific inhibitors in modified Whitten's medium for 24 h. Treatment with inhibitors that should inhibit myosin light chain kinase--HA100 (250 microM), Wortmannin (1 microM), and a combination of Wortmannin (1 microM), KT5720 (75 nM), and Iso-H7 (50 microM)--resulted in significantly higher pronuclear development (74.0%, 18.0%, and 35.0%, respectively) than in the negative control, H7 (10 microM; 2.0-12.4% depending upon the replication). Treatment with HA100 (250 microM) resulted in the dephosphorylation of the 25-kDa protein to a 22-kDa protein in 80.0% (n = 10) of oocytes exposed. However, Wortmannin (1 microM; n = 17), KT5720 (75 nM; n = 16), and Iso-H7 (50 microM; n = 19) treatment individually and in combination (n = 19) did not result in significant (p < 0.05; n = 19) dephosphorylation over the negative control, H7 (10 microM; n = 19). HA100 treatment resulted in significant cortical granule exocytosis when evaluated by laser confocal microscopy. In addition, protein kinase assays revealed lower myosin light chain kinase activity in electroactivated oocytes (p < 0.05) and protein kinase inhibitor-treated oocytes (p < 0.05) than in negative controls, nonelectroactivated oocytes, and H7 (10 microM)-treated oocytes. Treatment with HA100 (250 microM) resulted in pronuclear formation, dephosphorylation of the 25-kDa protein, and some release of cortical granules. These observations suggest that inhibition of myosin light chain kinase, protein kinase A, and protein kinase C results in activation of porcine oocytes.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗

Hemps, a novel EGF-like protein, plays a central role in ascidian metamorphosis.

All chordates share several characteristic features including a dorsal hollow neural tube, a notochord, a pharynx and an endostyle. Unlike other chordate taxa, ascidians have a biphasic life-history with two distinct body plans. During metamorphosis, the larval nerve cord and notochord degenerate and the pharyngeal gill slits and endostyle form. While ascidians, like other marine invertebrates, metamorphose in response to specific environmental cues, it remains unclear how these cues trigger metamorphosis. We have identified a novel gene (Hemps) which encodes a protein with a putative secretion signal sequence and four epidermal growth factor (EGF)-like repeats which is a key regulator of metamorphosis in the ascidian Herdmania curvata. Expression of Hemps increases markedly when the swimming tadpole larva becomes competent to undergo metamorphosis and then during the first 24 hours of metamorphosis. The Hemps protein is localised to the larval papillae and anterior epidermis of the larva in the region known to be required for metamorphosis. When the larva contacts an inductive cue the protein is released, spreading posteriorly and into the tunic as metamorphosis progresses. Metamorphosis is blocked by incubating larvae in anti-Hemps antibodies prior to the addition of the cue. Addition of recombinant Hemps protein to competent larvae induces metamorphosis in a concentration-dependent manner. A subgroup of genes are specifically induced during this process. These results demonstrate that the Hemps protein is a key regulator of ascidian metamorphosis and is distinct from previously described inducers of this process in terrestrial arthropods and aquatic vertebrates.

Animals↗

An unusual complication of T-tubes.

The morbidity of long-term ventilation tubes (Goode T-tubes) is often discussed with reference to otorrhoea, tympanosclerosis and long-term perforation. We report three cases of the T-tube slipping into the middle-ear cleft. In two of the three patients this was asymptomatic and the T-tubes were left in situ. In the third patient this complication was documented on three occasions when the T-tube was removed and reinserted for recurrent effusions. We are unaware of this complication being previously reported in the literature.

Adult↗

Multiple cutaneous metastases from laryngeal carcinoma.

A case of laryngeal squamous cell carcinoma recurrence presenting as multiple cutaneous metastases is presented. Such metastases are rare and are associated with a poor prognosis. Treatment is usually aimed at providing pain relief in these patients with a limited life expectancy.

Carcinoma, Squamous Cell↗

Systemic absorption of gentamicin ear drops.

Aminoglycoside-containing topical preparations are commonly used in the treatment of chronic suppurative otitis media and otitis externa. We report a case where systemic absorption of topical Gentisone HC occurred producing a serum gentamicin level of 6.2 micrograms/ml.

Absorption↗

Analyses of HLA-DQ alleles in recurrent spontaneous abortion (RSA) couples.

PROBLEM: The influence of HLA sharing on pregnancy outcome is controversial. In renal transplantation, HLA-DQB1 donor-recipient mismatches have been shown beneficial for long-term transplant success. Since pregnancy is defined as Nature's allograft, we investigated the relevance of HLA-DQ mismatching in normal reproducing couples compared to couples experiencing RSA. METHOD: Unexplained RSA couples referred to our laboratory for immunological testing were classified by immunological findings and obstetrical history. Primary RSA couples shared > or = 2 HLA-A, B, or DR antigens, had no cytotoxic anti-paternal antibodies, and no gestation beyond 20 weeks. Secondary RSA couples had cytotoxic anti-paternal antibodies and RSA after a live birth. HLA-DQA1 and DQB1 alleles were identified by PCR-SSP. RESULTS: No differences in DQA1 and DQB1 mismatch were observed among RSA patients and controls. DQA1-DQB1 haplotype mismatches were not different among the three groups of couples. CONCLUSIONS: In contrast to renal transplant, HLA-DQ incompatibility did not differ among RSA couples compared with successful reproducing couples.

Abortion, Habitual↗

An experimental vaccine cocktail for Plasmodium falciparum malaria.

Surface proteins from several different life-cycle stages of the malaria parasite Plasmodium falciparum were expressed at high levels in the yeast Saccharomyces cerevisiae. Purified proteins, both individually and in cocktails, were used to immunize mice and goats in conjunction with either Freund's adjuvant or a muramyl tripeptide-based adjuvant. Immune responses were measured by enzyme-linked immunosorbent assays and by the ability of antisera to inhibit (1) the invasion of hepatocytes by live sporozoites, (2) in vitro invasion of human erythrocytes by live merozoites, and (3) the development of oocytes in the mosquito vector. These results suggest that cocktails of different stage-specific antigens can provide the components necessary to block the development of the malaria parasite at multiple stages of its life cycle.

Animals↗

Recombinant Pfs25 protein of Plasmodium falciparum elicits malaria transmission-blocking immunity in experimental animals.

Pfs25 is a sexual stage antigen of Plasmodium falciparum that is expressed on the surface of zygote and ookinete forms of the parasite. Monoclonal antibodies directed against native Pfs25 can block completely the development of P. falciparum oocysts in the midgut of the mosquito vector. Thus, this 25-kD protein is a potential vaccine candidate for eliciting transmission-blocking immunity in inhabitants of malaria endemic regions. We have synthesized, by secretion from yeast, a polypeptide analogue of Pfs25 that reacts with conformation-dependent monoclonal antibodies, and elicits transmission-blocking antibodies when used to immunize mice and monkeys in conjunction with a muramyl tripeptide adjuvant. Our results suggest the further evaluation of recombinant DNA-derived Pfs25 in transmission-blocking vaccination studies in humans.

Animals↗

Immunogenicity of recombinant Plasmodium falciparum SERA proteins in rodents.

We have expressed defined regions of the serine-repeat antigen (SERA) of the Honduras-1 strain of Plasmodium falciparum in the yeast Saccharomyces cerevisiae. Amino-terminal domains of the natural SERA protein have been shown previously to be targets for parasite-inhibitory murine monoclonal antibodies. Two recombinant SERA antigens were selected for purification and immunological analysis. The first (SERA 1), corresponding to amino acids 24-285 of the natural SERA precursor, was expressed by the ubiquitin fusion method. This allowed for in vivo cleavage by endogenous yeast ubiquitin hydrolase, and subsequent isolation of the mature polypeptide. The second, larger protein (SERA N), encompassing amino acids 24-506, was expressed at only low levels using this system, but could be isolated in high yields when fused to human gamma-interferon (gamma-IFN). Each purified protein was used to immunize mice with either Freund's adjuvant or a muramyl tripeptide adjuvant that has been used in humans. Sera from immunized mice were shown to be capable of in vitro inhibition of invasion of erythrocytes by the Honduras-1 strain of P. falciparum. The results suggest that a recombinant SERA antigen may be an effective component of a candidate malaria vaccine.

Animals↗

Protective immunity induced in Aotus monkeys by recombinant SERA proteins of Plasmodium falciparum.

We describe the vaccination of Panamanian monkeys (Aotus sp.) with two recombinant blood stage antigens that each contain a portion of the N-terminal region of the SERA (serine repeat antigen) protein of the malaria parasite Plasmodium falciparum. We immunized with either a 262-amino-acid SERA fragment (SERA I) that contains amino acids 24 to 285 of the 989-amino-acid protein or a 483-amino-acid SERA fragment (SERA N) that contains amino acids 24 to 506 as part of a fusion protein with human gamma interferon. The recombinant proteins were shown to stimulate protective immunity when administered with complete and incomplete Freund adjuvant. Four of six immunized monkeys challenged by intravenous inoculation with blood stage P. falciparum developed parasitemias that were reduced by at least 1,000-fold. Two of six immunized monkeys developed parasitemias which were comparable to the lowest parasitemia in one of four controls and were 50- to 1,000-fold lower than in the other three controls.

Animals↗