Laboratory tests in uveitis - New developments in the analysis of local antibody production

Analysis of local intraocular antibody production is a valuable tool with which to confirm a suspected clinical diagnosis in uveitis. We have analysed paired serum and aqueous samples for the presence of specific antibodies against toxoplasma, cytomegalovirus, herpes simplex virus and varicella zoster virus. Of the patients retrospectively diagnosed as having toxoplasma chorioretinitis 75% had a positive antibody coefficient indicating specific antibody production in the eye. Local antibody production in the eye directed against CMV confirmed the suspected diagnosis of CMV retinitis in 50% of the AIDS patients investigated. So far we have not been able to demonstrate local antibody production against herpes simplex virus (26 samples tested). Two of three patients with acute retinal necrosis had a positive antibody coefficient against varicella zoster virus. Both of these patients had an even higher titer in the aqueous than in serum. Since the choice of therapy, in infectious uveitis, depends on the causative organisms, it is very important to confirm a suspected clinical diagnosis by means of aqueous humor analysi

The last month of Szent-Gyorgyi in Groningen

Albert (von) Szent-Gyorgyi started his studies on biological oxidation processes - which also resulted in the discovery of vitamin C, for which he received the Nobel Price in 1937 - in the Laboratory of Physiology of the University in Groningen in 1922-1926. These studies were later continued in Cambridge (UK) and Szeged (Hungary). When he had already received the invitation as well as the financial means to come and work in Cambridge, he still did experiments in Groningen to find out whether the adrenal extract, isolated by him and later found to be a major source of vitamin C, contained the hormone essential for the survival of cats whose adrenals were removed. He was rather upset by the negative results of this experiment, judging by the recollections of a former student of his. This history constitutes an interesting example of the difference between serendipitous discovery and planned invention

Do asparagine-linked carbohydrate chains in glycoproteins have a preference for beta-bends?

X-ray structures of the conformation of carbohydrate moieties and connected regions of glycoproteins are summarized. Evidence is presented that there is some preference for carbohydrate attachment at β-bends. Evolution may have favored glycosylation to occur at bends to ensure free mobility of the carbohydrate moieties

Selection Against Glycosylation in Ruminant Pancreatic Ribonucleases by Replacements in the Ancestral Carbohydrate Attachment Site

A hypothesis, proposed 25 years ago, that there is selection against glycosylation in ruminant pancreatic ribonucleases by replacement of methionine to leucine in the ancestral carbohydrate attachment site Asn–Met–Thr at residues 34–36, was experimentally confirmed. The replacement of leucine at position 35 by methionine in bovine ribonuclease resulted in a three-fold relative increase in glycosylation when expressed in Chinese hamster ovary cells.ISSN:0006-2928ISSN:1573-492

The primary structures of pancreatic ribonucleases from African porcupine and casiragua, two hystricomorph rodent species

The amino acid sequences of the pancreatic ribonucleases from African porcupine (Hystrix cristata) and casiragua (Proechimys guairae, a caviomorph rodent species related to the coypu) were determined. The ribonucleases were isolated form minces of pancreatic tissue which had been used for the extraction of the insulins. The results of the sequence determinations of residues 67–78 in both enzymes were ambiguous. Therefore, homology with other ribonucleases has been used in deriving these sequences. At position 94 aspartic acid was found, while ali other ribonuclease sequenced to date have asparagine at this position. This may indicate a specific deamidation as a result of the acidic conditions during the extraction of insulin. The amino acid sequence of African porcupine ribonuclease shows a close relationship with those of the South-American caviomorph rodents, which implies that the hystricomorph suborder of the rodents, to which both the African porcupine and the caviomorphs belong, is a natural (evolutionary) taxon. Both porcupine and casiragua ribonuclease are glycoproteins with complex-type carbohydrate chains attached to asparagine34

The amino acid sequence of snapping turtle (Chelydra serpentina) ribonuclease

Snapping turtle (Chelydra serpentina) ribonuclease was isolated from pancreatic tissue. Turtle ribonuclease binds much more weakly to the affinity chromatography matrix used than mammalian ribonucleases. The amino acid sequence was determined from overlapping peptides obtained from three different digests. The N-terminal amino acid sequence of the protein determined by others and homology were used as additional evidence for several overlaps. The polypeptide chain consists of 119 amino acid residues. Compared to most ribonucleases the N-terminal residue, three residues in the loop near residue 71 and two residues in the loop near residue 114 are deleted, and there is one additional residue in the loop near residue 23. The half-cystines at positions 65 and 72, which form a disulfide bond in mammalian ribonucleases, are not present in turtle ribonuclease. Turtle ribonuclease differs from bovine ribonuclease at 70 of the 118 positions where both proteins have amino acid residues. Turtle ribonuclease contains no carbohydrate, although the enzyme possesses a recognition site for carbohydrate attachment in the sequence Asn-Ala-Ser (positions 76-78).