Influence of V5/6-His Tag on the Properties of Gap Junction Channels Composed of Connexin43, Connexin40 or Connexin45

HeLa cells expressing wild-type connexin43, connexin40 or connexin45 and connexins fused with a V5/6-His tag to the carboxyl terminus (CT) domain (Cx43-tag, Cx40-tag, Cx45-tag) were used to study connexin expression and the electrical properties of gap junction channels. Immunoblots and immunolabeling indicated that tagged connexins are synthesized and targeted to gap junctions in a similar manner to their wild-type counterparts. Voltage-clamp experiments on cell pairs revealed that tagged connexins form functional channels. Comparison of multichannel and single-channel conductances indicates that tagging reduces the number of operational channels, implying interference with hemichannel trafficking, docking and/or channel opening. Tagging provoked connexin-specific effects on multichannel and single-channel properties. The Cx43-tag was most affected and the Cx45-tag, least. The modifications included (1) Vj-sensitive gating of Ij (Vj, gap junction voltage; Ij, gap junction current), (2) contribution and (3) kinetics of Ij deactivation and (4) single-channel conductance. The first three reflect alterations of fast Vj gating. Hence, they may be caused by structural and/or electrical changes on the CT that interact with domains of the amino terminus and cytoplasmic loop. The fourth reflects alterations of the ion-conducting pathway. Conceivably, mutations at sites remote from the channel pore, e.g., 6-His-tagged CT, affect protein conformation and thus modify channel properties indirectly. Hence, V5/6-His tagging of connexins is a useful tool for expression studies in vivo. However, it should not be ignored that it introduces connexin-dependent changes in both expression level and electrophysiological properties

Does Landscape Fragmentation Influence Sex Ratio of Dioecious Plants? A Case Study of Pistacia chinensis in the Thousand-Island Lake Region of China

The Thousand-Island Lake region in Zhejiang Province, China is a highly fragmented landscape with a clear point-in-time of fragmentation as a result of flooding to form the reservoir. Islands in the artificial lake were surveyed to examine how population sex ratio of a dioecious plant specie Pistacia chinensis B. was affected by landscape fragmentation. A natural population on the mainland near the lake was also surveyed for comparison. Population size, sex ratio and diameter at breast height (DBH) of individuals were measured over 2 years. More than 1,500 individuals, distributed in 31 populations, were studied. Soil nitrogen in the different populations was measured to identify the relationship between sex ratio and micro-environmental conditions. In accordance with the results of many other reports on biased sex ratio in relation to environmental gradient, we found that poor soil nitrogen areas fostered male-biased populations. In addition, the degree of sex ratio bias increased with decreasing population size and population connectivity. The biased sex ratios were only found in younger individuals (less than 50 years old) in small populations, while a stable 1∶1 sex ratio was found in the large population on the mainland. We concluded that the effects of landscape fragmentation on the dioecious population sex ratio were mainly achieved in relation to changing soil nitrogen conditions in patches and pollen limitation within and among populations. Large populations could maintain a more suitable environment in terms of nutrient conditions and pollen flow, subsequently maintaining a stable sex ratio in dioecious plant populations. Both micro-environmental factors and spatial structure should be considered in fragmented landscape for the conservation of dioecious plant species

Cirurgia de osteoma de seio frontal Surgery of frontal sinus osteoma

Os osteomas do seio frontal correspondem a 57% dos osteomas dos seios paranasais, com incidência variando de 0,01% a 3%. A remoção cirúrgica nos osteomas frontais é indicada nos pacientes sintomáticos. Nos pacientes assintomáticos pode-se adotar a conduta conservadora ou cirúrgica em todos os pacientes independente da sua localização ou extensão. Cinco pacientes com diagnóstico de osteoma de seio frontal foram operados entre 1995 e 1999. A idade média foi 38,4 anos (extremos de 12 a 55 anos), sendo 3 homens e 2 mulheres. O período de sintomatologia variou de 6 meses a 3 anos com média de 10,5 meses. Quatro pacientes apresentaram cefaléia. Um paciente apresentou epistaxe. Os exames complementares realizados foram: radiografia simples e tomografia computadorizada de seios paranasais com cortes axiais e coronais. Em dois pacientes o diâmetro do osteoma foi maior que 3 cm, e menor que 3 cm em três. A decisão da técnica cirúrgica entre coronal e supraciliar foi estética, reservando-se a abordagem supraciliar para um paciente com calvície, apesar do tumor ser volumoso com extensão para seio etmoidal. Nenhuma dificuldade técnica intra-operatória foi atribuída à escolha da abordagem. O óstio nasofrontal não foi obstruído no intra-operatório. O seguimento pós-operatório mínimo foi de dois anos. Em todos os casos a remoção foi total sem recidiva ou resíduos tumorais. Os sintomas clínicos, achados radiológicos e abordagens cirúrgicas são discutidos. Não ocorreram complicações pós-operatórias.<br>Frontal sinus osteomas are 57% of all paranasal sinus osteomas, with an incidence of 00.1 to 3%. Surgical removal of the frontal sinus osteomas is done in symptomatic patients. Asymptomatic patients can be managed conservatively or submitted to surgery in spite of its location or extension. Five patients having the diagnosis of frontal sinus osteoma were operated on between 1995 and 1999. Medium age was 38.4 years (from 12 to 55 years), 3 male and 2 female. Symptoms occurred from 6 months to 3 years, average of 10.5 months. Four patients had previous headache and one had epistaxis. All patients had standard radiological exams and computed tomography with coronal and axial studies of paranasal sinus. In two patients the diameter of the osteoma was larger than 3 cm and in three smaller than 3 cm. The choice between coronal and supraciliar approach was made according to esthetics, supraciliar approach was made in only one bald patient even with the tumor being large and extending to ethmoidal sinus. Any intra operative difficulty was related to the choice of the approach. Naso-frontal ostium was not obstructed in intra operative course. Minimal postoperative follow up was of two years. Osteomas were radically removed in all patients with no recurrence or residual tumor. Clinical findings, radiological exams and surgical approaches are discussed. No postoperative complications occurred