Cloning of Multiple Forms of Goldfish Vimentin: Differential Expression in CNS

Abstract: In efforts to determine the primary structure of intermediate filament proteins in the goldfish visual pathway, we isolated clones from a retinal λgt11 cDNA expression library that represent goldfish vimentin. We show that there are at least two forms of goldfish vimentin, designated as vimentin α and vimentin β. RNase protection assays indicate that vimentin α mRNA is expressed in low amounts in retina, optic nerve, and brain and in higher amounts in spinal cord. In contrast, vimentin β mRNA is expressed in low amounts in retina, optic nerve, brain, and spinal cord and in very high amounts in eye lens. Immunohistochemical studies show that in the optic nerve, vimentin α is mainly restricted to blood vessels, meninges, and septa. Light staining is observed with this antibody in an astrocytic glial pattern throughout the optic nerve. Two-dimensional gel analysis shows that all of these goldfish vimentins are low abundant components of optic nerve cytoskeletal preparations.     

SINGLE-CELL PIGMENTATION OF PORPHYRA LINEARIS ANALYZED BY FOURIER TRANSFORM MULTI-PIXEL SPECTROSCOPY AND IMAGE ANALYSIS1

The novel method of Fourier transform multi-pixel spectroscopy was used for the nondestructive analysis of and comparison of pigmentation in different regions of live thalli of the red alga Porphyra linearis. Because the thallus in this alga consists of a monolayer of nonoverlapping cells, we were able to analyze the pigmentation of single cells by combining light absorbance with natural fluorescence data. From the image of each cell in the vegetative male and female reproductive and holdfast regions, more than 4 ± 10⁴ fluorescence and absorbance spectra were obtained. Specific pigments in the different regions were localized by the use of a software program of similarity mapping followed by image construction. The reconstructed images revealed subcellular localization of each pigment according to specific spectroscopic fingerprints. The results showed that the vegetative and female reproductive cell types had a significantly higher content of phycoerythrin than of phycocyanin, and quite similar chlorophyll a levels. Most of the holdfast cells were poorly pigmented, but had more chlorophyll a than phycoerythrin or phycocyanin. The male reproductive cells contained only traces of pigments. Thus, by using Fourier transform multipixel spectroscopy, we were able to characterize the pigmentation of different regions of the thallus and follow the distribution patterns of the different pigments on the subcellular level along the differentiation gradient of the alga.     

A single nucleotide substitution in TaHKT1;5-D controls shoot Na+ accumulation in bread wheat

Improving salinity tolerance in the most widely cultivated cereal, bread wheat (Triticum aestivum L.), is essential to increase grain yields on saline agricultural lands. A Portuguese landrace, Mocho de Espiga Branca accumulates up to sixfold greater leaf and sheath sodium (Na⁺) than two Australian cultivars, Gladius and Scout, under salt stress in hydroponics. Despite high leaf and sheath Na⁺ concentrations, Mocho de Espiga Branca maintained similar salinity tolerance compared to Gladius and Scout. A naturally occurring single nucleotide substitution was identified in the gene encoding a major Na⁺ transporter TaHKT1;5-D in Mocho de Espiga Branca, which resulted in a L190P amino acid residue variation. This variant prevents Mocho de Espiga Branca from retrieving Na⁺ from the root xylem leading to a high shoot Na⁺ concentration. The identification of the tissue-tolerant Mocho de Espiga Branca will accelerate the development of more elite salt-tolerant bread wheat cultivars.     

Identification of a streptococcal octapeptide motif involved in acute rheumatic fever

Acute rheumatic fever is a serious autoimmune sequela of pharyngitis caused by certain group A streptococci. One mechanism applied by streptococcal strains capable of causing acute rheumatic fever is formation of an autoantigenic complex with human collagen IV. In some geographic regions with a high incidence of acute rheumatic fever pharyngeal carriage of group C and group G streptococci prevails. Examination of such strains revealed the presence of M-like surface proteins that bind human collagen. Using a peptide array and recombinant proteins with targeted amino acid substitutions, we could demonstrate that formation of collagen complexes during streptococcal infections depends on an octapeptide motif, which is present in collagen binding M and M-like proteins of different beta-hemolytic streptococcal species. Mice immunized with streptococcal proteins that contain the collagen binding octapeptide motif developed high serum titers of anti-collagen antibodies. In sera of rheumatic fever patients such a collagen autoimmune response was accompanied by specific reactivity against the collagen-binding proteins, linking the observed effect to clinical cases. Taken together, the data demonstrate that the identified octapeptide motif through its action on collagen plays a crucial role in the pathogenesis of rheumatic fever. Eradication of streptococci that express proteins with the collagen binding motif appears advisable for controlling rheumatic fever.     

Group G streptococcal IgG binding molecules FOG and protein G have different impacts on opsonization by C1q

Recent epidemiological data on diseases caused by beta-hemolytic streptococci belonging to Lancefield group C and G (GCS, GGS) underline that they are an emerging threat to human health. Among various virulence factors expressed by GCS and GGS isolates from human infections, M and M-like proteins are considered important because of their anti-phagocytic activity. In addition, protein G has been implicated in the accumulation of IgG on the bacterial surface through non-immune binding. The function of this interaction, however, is still unknown. Using isogenic mutants lacking protein G or the M-like protein FOG (group G streptococci), respectively, we could show that FOG contributes substantially to IgG binding. A detailed characterization of the interaction between IgG and FOG revealed its ability to bind the Fc region of human IgG and its binding to the subclasses IgG1, IgG2, and IgG4. FOG was also found to bind IgG of several animal species. Surface plasmon resonance measurements indicate a high affinity to human IgG with a dissociation constant of 2.4 pm. The binding site was localized in a central motif of FOG. It has long been speculated about anti-opsonic functions of streptococcal Fc-binding proteins. The presented data for the first time provide evidence and, furthermore, indicate functional differences between protein G and FOG. By obstructing the interaction between IgG and C1q, protein G prevented recognition by the classical pathway of the complement system. In contrast, IgG that was bound to FOG remained capable of binding C1q, an effect that may have important consequences in the pathogenesis of GGS infections.     

The role of static features of males in the mate choice behavior of female Japanese quail (Coturnix japonica)

In two experiments, we investigated the mate choice behavior of female Japanese quail toward taxidermically-prepared male models. Both experiments consisted of four phases: (1) habituation; (2) a pre-test in which two taxidermically-prepared models of male birds were presented; (3) observation in which the respective non-preferred male model was presented either alone or with another stimulus, and (4) a post-test in which male models were again presented alone. Results showed that focal females increased their preference for a non-preferred male model that they had previously observed with a live female (Experiment 1) or with a taxidermically-prepared female model (Experiment 2). Two control groups ruled out the possibility that focal females were choosing male models either because: (1) males were presented with an additional stimulus, or (2) females were choosing an area where they observed male models with other females. The findings suggest that female quail may utilize static, species-specific features of male conspecifics in mate choice.     

The long-term results of ruby laser depilation in a consecutive series of 346 patients

The goal of this study was to prospectively assess the long-term results of ruby laser depilation in 346 consecutive patients who underwent hair removal at 402 anatomical sites. The patients were treated using a ruby laser, with mean power ranging from 8.6 J to 15.7 J according to skin type. Results were assessed using two outcome measures-the percentage reduction in hair density and the hair-free interval. The median reduction in hair density was 55 percent (range, 0 to 100 percent) at a median time of 1 year after the last treatment session. The median hair-free interval was 8 weeks. Patients underwent a median number of four treatment sessions. Forty-three of the 346 patients were treated at more than one anatomical site. Of the sites treated, 75 percent reduction in hair density was achieved in 22 percent, 90 percent reduction was achieved in 2.2 percent, and complete depilation was achieved in only 0.7 percent. Darker colored hair was more effectively treated. Treatment efficacy was not affected by anatomical site, with the exception of the faces of male patients, which were found to be particularly resistant to treatment. There was a significant correlation between the number of treatments given and the outcome. The overall complication rate was 9.0 percent (36 of 402 sites) with respect to pigmentary changes and blistering, but varied according to Fitzpatrick skin type. The complication rate was highest in skin types V and VI (24.7 percent), with no complications in skin type I. Although a greater than 50 percent reduction in hair density was achieved in half of the 346 patients treated, complete depilation was achieved in only an extremely limited number of patients.     

A new method of subcutaneous placement of free jejunal flaps to reconstruct a diversionary conduit for swallowing in complicated pharyngoesophageal injury

Choking is a serious problem in pharyngoesophageal reconstruction, which may occur following tumor ablation of the pharynx or following corrosive injury involving the epiglottis and other parts of the upper airway. To prevent choking and the risk of severe pulmonary complications, patients have to give up oral intake and assume feeding via jejunostomy for the rest of their lives. After reconstruction of the esophagus, eight patients experienced frequent choking and aspiration. With a free jejunal flap, the inlet for food could be separated from the route of the upper airway by a diversion technique. The jejunum segment was transferred microsurgically to reconstruct the cervical esophagus, with its inlet at the buccogingival sulcus. There were no surgical complications related to either the free jejunal flap transfer or the donor site. Postoperatively, patients require re-education of their pattern of swallowing, but after the rehabilitation period all patients reported a satisfactory oral intake through the reconstructed esophagus to the abdomen without choking. There were no episodes of aspiration following reconstruction. With this new method to create a separate food pathway, patients can resume oral intake safely without choking and without permanent jejunostomy. This technique offers a useful solution for patients who suffer from recurrent choking and aspiration following injury or ablation of the pharynx.     

Osteotomy to treat malocclusion following reconstruction of the mandible with the free fibula flap

Malocclusion may result after free fibula flap reconstruction of the mandible, because of inadequate positioning of the temporomandibular joint, inaccurate contouring of the reconstruction plate, or subsequent fracture of a miniplate. Factors that alter the vascularity of the transplanted fibula may also result in a delayed presentation of malocclusion. Seven cases are presented, in which primary surgical treatment consisted of segmental mandibulectomy and reconstruction with a free fibula osteoseptocutaneous flap. Fixation was achieved with a reconstruction plate in five cases and a miniplate in two cases. Malocclusion was corrected with an osteotomy performed at the junction of the fibula and the native mandible. The new osteotomy sites were fixed with miniplates and maintained with intermaxillary fixation. Complete bony union was achieved at the osteotomy sites. The correction of malocclusion was successful in all cases, and all patients have resumed a normal diet. This report demonstrates that osteotomy and realignment of the mandible are effective for the secondary correction of malocclusion after mandibular reconstruction with the free fibula osteoseptocutaneous flap.