Expression and characterization of human lamin C

We have expressed human lamin C cDNA in E. coli using a modification of the pLcII vector system. Protein produced in this way had seven additional amino acids at its N-terminus, but retained key lamin structural and assembly properties. The modified vector we produced may prove useful when difficulties are encountered in removal of the cII fusion peptide by factor X cleavage in the pLcII system. Shadowed preparations of expressed lamin C showed the presence of 50-nm rod-like particles that closely resembled those observed for native material. Isolated molecules had two globular domains at one end, indicating that they were constructed from two parallel polypeptide chains. The expressed material also formed paracrystals with a characteristic 22.5 nm axial repeat, indicating that its assembly properties had also been retained. We also used site-specific mutagenesis to engineer a lamin fragment that lacked the C-terminal non-helical domain of the molecule. This material formed paracrystals similar to those obtained with the intact molecule, indicating that the large C-terminal non-helical domain did not contain information vital for lamin assembly.     

Neuropeptide regulation of human dermal microvascular endothelial cell ICAM-1 expression and function

There isincreasing evidence that sensory nerves may participate in cutaneousinflammatory responses by the release of neuropeptides such assubstance P (SP). We examined the direct effect of SP on human dermalmicrovascular endothelial cell (HDMEC) intercellular adhesion molecule1 (ICAM-1) expression and function. Our results indicated that,although cultured HDMEC expressed mRNA for neurokinin receptors 1, 2, and 3 (NK-1R, NK-2R, and NK-3R), SP initiated a rapid increase in HDMECintracellular Ca²⁺ levels,primarily by the activation of NK-1R. Immunohistochemistry studieslikewise demonstrated that HDMEC predominantly expressed NK-1R. Theaddition of SP to HDMEC resulted in a rapid increase in cellular ICAM-1mRNA levels, followed by a fivefold increase in ICAM-1 cell surfaceexpression. This functionally resulted in a threefold increase in⁵¹Cr-labeled binding of J-Ylymphoblastoid cells to HDMEC. In vivo studies demonstrated a markedincrease in microvascular ICAM-1 immunostaining 24 and 48 h afterapplication of capsaicin to the skin. These results indicate thatneuropeptides such as SP are capable of directly activating HDMEC toexpress increased levels of functional ICAM-1 and further support therole of the cutaneous neurological system in modulating inflammatoryprocesses in the skin.

     

Clinical Evaluation of Perhexiline Maleate in Patients with Angina Pectoris

This paper reports a double-blind trial of a new antianginal drug, perhexiline. Fifty-five patients suffering from angina pectoris were studied for periods of 12 or 24 weeks in a cross-over comparison against a placebo in four centres in the United Kingdom and Ireland. Perhexiline was effective in most patients as judged by reducing the number of anginal attacks in 84% and the consumption of glyceryl trinitrate tablets in 64%. The major side effect, dizziness, noted in one-third of the patients, may be dose/body-weight related. Perhexiline is a valuable new agent for the treatment of patients with angina, especially those who do not respond to other antianginal agents.     

Modification of Pattern of Photosynthate Movement within and between Shoots of Vitis vinifera L

During the prebloom and bloom stages, no movement of labeled photosynthates occurred from a shoot of Vitis vinifera L. fed with ¹⁴CO₂, to an adjacent shoot on the same spur. Movement of labeled assimilates into the unfed shoot was induced when this shoot was sprayed with 2.89 × 10⁻³m gibberellic acid during the prebloom stage. During the bloom stage darkening or defoliation of the unfed shoot resulted in the import of labeled photosynthates from the adjacent fed shoot. Similarly, movement of ¹⁴C into an untreated shoot was induced by removing the terminal 7.5 centimeters and deblossoming the fed shoot. During the berry set stage, translocation of labeled photosynthates from a newly exporting leaf was upwards to the shoot tip, but the direction of movement was reversed by removal of the shoot tip or by darkening or removal of the leaves below the fed leaf. Translocation of photosynthates was predominantly basipetal from a fed leaf near the base of the shoot during the berry set stage, but upward movement was induced by darkening or defoliation of the upper part of the shoot.     

The Ulcerating Tuberculous Hilar Gland

In primary infection tuberculosis, the infected hilar gland(s) may cause involvement of peripheral lung tissue not only by pressure but also by rupture and discharge of caseous material into a bronchus. Atelectasis or lung infection or both may result and bronchiectasis may ensue.Early bronchoscopy is required when this form of tuberculosis fails to subside promptly under treatment.Bronchography is indicated to detect residual bronchiectasis which should be removed surgically.Three of six proved cases of Group A tuberculous tracheobronchitis caused by an ulcerating hilar gland required pulmonary resection for removal of residual bronchiectasis; two of these were complicated by atelectasis. All six patients are alive and well.     

Decreased snake venom metalloproteinase effects via inhibition of enzyme and modification of fibrinogen

Since the introduction of antivenom administration 120 years ago to treat venomous snake bit, it has been the gold standard for saving life and limb. However, this therapeutic approach is not always effective and not without potential life-threatening side effects. We tested a new paradigm to abrogate the plasmatic anticoagulant effects of fibrinogenolytic snake venom metalloproteinases by modification of fibrinogen with iron and carbon monoxide and by inhibiting these Zn²⁺ dependent metalloproteinases directly with carbon monoxide exposure. Assessment of the fibrinogenolytic effects of venoms collected from Puff adder, Gaboon viper and Indian cobra snakes on plasmatic coagulation kinetics was performed with thrombelastography. Pretreatment of plasma with iron and carbon monoxide exposure markedly attenuated the effects of all three venoms, and direct pretreatment of each venom with carbon monoxide also significantly decreased the ability to compromise coagulation. These results demonstrated that the introduction of a transition metal (e.g., modulation of the α-chain of fibrinogen with iron), modulation of transition metal in heme (e.g., carbon monoxide modulation of fibrinogen-bound heme iron), and direct inhibition of transition metal containing venom enzymes (e.g., CO binding to Zn²⁺ or displacing Zn²⁺ from the catalytic site) significantly decreased fibrinogenolytic activity. This biometal modulation strategy to attenuate the anticoagulant effects of snake venom metalloproteinases could potentially diminish hemostatic injury in envenomed patients until antivenom can be administered.