Co-localization of components of the protein-synthesizing machinery with the cytoskeleton in G0-arrested cells

The distribution of eukaryotic elongation factor 2 (eEF-2) in G0-arrested fixed human skin diploid fibroblasts was studied by indirect immunofluorescent microscopy. It was found earlier that the main part of eEF-2 in cycling cells was located near the nucleus in the endoplasm (Gavrilova et al., 1987). It has been demonstrated here that the transition from proliferation to the G0 phase of the cell cycle leads to the distribution of eEF-2 mainly along the intermediate filaments and/or microtubules. Both in cycling and in G0-arrested fibroblasts a portion of eEF-2 is also co-localized with actin microfilament bundles. The reversion of the cells from the G0 phase to proliferation is accompanied by rearrangement of the actin cytoskeleton and reversal to the original pattern of eEF-2 distribution. It is likely that the different types of cytoskeleton in eukaryotic cells can be involved in organization of protein-synthesizing machinery.     

Partitioning of the Golgi apparatus during mitosis in living HeLa cells

The Golgi apparatus of HeLa cells was fluorescently tagged with a green fluorescent protein (GFP), localized by attachment to the NH2-terminal retention signal of N-acetylglucosaminyltransferase I (NAGT I). The location was confirmed by immunogold and immunofluorescence microscopy using a variety of Golgi markers. The behavior of the fluorescent Golgi marker was observed in fixed and living mitotic cells using confocal microscopy. By metaphase, cells contained a constant number of Golgi fragments dispersed throughout the cytoplasm. Conventional and cryoimmunoelectron microscopy showed that the NAGT I-GFP chimera (NAGFP)-positive fragments were tubulo-vesicular mitotic Golgi clusters. Mitotic conversion of Golgi stacks into mitotic clusters had surprisingly little effect on the polarity of Golgi membrane markers at the level of fluorescence microscopy. In living cells, there was little self-directed movement of the clusters in the period from metaphase to early telophase. In late telophase, the Golgi ribbon began to be reformed by a dynamic process of congregation and tubulation of the newly inherited Golgi fragments. The accuracy of partitioning the NAGFP-tagged Golgi was found to exceed that expected for a stochastic partitioning process. The results provide direct evidence for mitotic clusters as the unit of partitioning and suggest that precise regulation of the number, position, and compartmentation of mitotic membranes is a critical feature for the ordered inheritance of the Golgi apparatus.     

Detection of P-glycoprotein in the Golgi apparatus of drug-untreated human melanoma cells

The intracellular location of the MDR1 gene product, known as P-glycoprotein (P-gp), has been detected by flow cytometry in 3 stabilized human melanoma cell lines which had never undergone cytotoxic drug treatment and did not express P-gp on the plasma membrane. In addition, MDR1 mRNA expression was revealed by RT-PCR in the same cell lines. Immunofluorescence microscopy, performed by using the same 2 monoclonal antibodies (MM4.17 and MRK-16) as employed in the flow-cytometric analysis, revealed the presence of P-gp intracytoplasmically, in a well-defined perinuclear region. Double immunofluorescence labelling and immunoelectron microscopy strongly suggested the location of the transporter molecule in the Golgi apparatus. The same observations have been obtained on a primary culture from a metastasis of human melanoma. Analysis of the expression of another membrane transport protein, the multidrug-resistance-related protein (MRP1), showed that it was present in the cytoplasm of all the melanoma cell lines examined. MRP1 also showed Golgi-like localization. The study by laser scanning confocal microscopy on the intracellular localization of the anti-tumoral agent doxorubicin (DOX) during the drug-uptake and -efflux phases, indicated the Golgi apparatus as a preferential accumulation site for the anthracyclinic antibiotic. P-gp function modulators (verapamil and cyclosporin A) were able to modify DOX intracytoplasmic distribution and to increase drug intracellular concentration and cytotoxic effect in melanoma cells. On the contrary, MRP1 modulators (probenecid and genistein) did not significantly influence either DOX efflux and distribution or the sensitivity of melanoma cells to the cytotoxic drug.     

Ribosomal apparatus of liver cells in the process of carinogenesis induced by 4-dimethylaminoazobenzene

The antigen of hepatitis B was repeatedly examined in 1356 adults patients with acute viral hepatitis. In 701 patients the HBAg was positive at the beginning of illness and in 657 was negative. The maximum of the negative HBAg results could be found in the lower age groups, whereas the maximum of the positive HBAg results was shifted to the 5th and 6th decade. In HBAg positive cases of hepatitis more numerous anicteric and asymptomatic forms could be observed on the other hand, however, at the same time also o more cholestatic forms and subacute necrotizing and malignant forms appeared. The HBAg positive hepatitis had on average also a more serious form and course. From the epidemiological point of view the admission to the hospital, which preceded in 51% of patients appeared as the most important factor. In 16.3% of patients the HBAg postivity persisted still 6 months after the discharge from the hospital.     

A model of the structure of the receptor apparatus of HIV target cells (1)

A model is proposed and well based of structure of the receptor apparatus of interacting HIV target cells: Langerhans cells (LC) and helper cells (HC)--designed to explain feasibility of realizing a double activation signal, together with some features of immune pathogenesis of HIV-infection (morphofunctional abnormalities in certain subpopulations of the immune system such as HC, suppressor cells, killer cells, natural killers, lymph cells, macrophages, LC), as well as immunogenetic predisposition to AIDS.     

Human breast cancer cells contain an error-prone DNA replication apparatus

The mechanisms responsible for creating genetic errors and genomic instability in cancer cells have not been fully defined. Recently, it has been shown that human cells contain a highly organized complex of proteins, termed the DNA synthesome, that is fully competent to carry out all phases of SV40 in vitro DNA replication (J. M. Coll et al, Oncol. Res., 8: 435-447, 1996; L. H. Malkas et al., Biochemistry, 29: 6362-6374, 1990; Y. Wu et al., J. Cell. Biochem., 54: 32-46, 1994; N. Applegren et al., J. Cell. Biochem., 54: 32-46, 1994). DNA replication fidelity analyses of the DNA synthesome derived from malignant and nonmalignant human breast cells demonstrate that the malignant cell synthesome is mutagenic. The decrease in tumor cell replication fidelity was not due to an increased proliferative capacity of the tumor cells or an increase in the synthetic activity of their DNA synthesome. The ratios of insertions, deletions, and mismatches created by the synthesome from malignant and nonmalignant breast cells were essentially identical, despite the greater overall number of mutations made by the breast cancer cell synthesome. These data define, for the first time, a mechanism unique to cancer cells that contributes to the observed increase in genetic mutation in cancer cells.     

Structure of the Golgi apparatus in stimulated and nonstimulated acinar cells of mammary glands of the rat

The structural features of the Golgi apparatus of acinar cells of mammary glands were examined with the electron microscope in 3 groups of rats: (1) in lactating female animals at 8 days postpartum, which served as controls; (2) in female rats sacrificed at various intervals from 2 to 30 hours following separation from their 8-day old pups; and (3) in females separated from their 8-day-old pups for a period of 12 hours and returned to their litters for durations of 1, 2, 4, and 8 hours. In animals of group 2, the Golgi stacks remained identical to that of controls between 2 and 8 hours. At 12 hours and later, the Golgi stacks decreased progressively in size, but the number of elements composing the stacks remained similar to that of lactating females and all contained casein submicelles. At 24 and 30 hours, typical secretory granules containing casein micelles disappeared from the trans aspect of the stacks. The earliest and most striking changes observed in the Golgi apparatus of the rats of group 2 took place at 12 hours. At this time, the prosecretory and secretory granules decreased considerably in volume and lost most of their electron-lucent content. This indicated that the delivery of small molecules, i.e., lactose and H2O, to these structures was soon altered following arrest of the sucking stimulus. In animals of group 3, the size of prosecretory and secretory granules and the amount of their electron-lucent content reverted to normal at 4 hours. Thus the influx of lactose and H2O into these structures appears to be rapidly restored after returning the pups to their mothers. The decrease in size of the Golgi stacks noted at 12, 18, and 24 hours following arrest of lactation (group 2), was accompanied by an increase in number of small vesicles that formed clusters next to the Golgi stacks and in "wells." Thus in these regressing Golgi stacks, many of the associated small vesicles appear to arise by vesiculation of the saccules.     

The regulation of calcium homeostasis in nerve cells

We have reviewed the major cellular elements related to the release and buffering of calcium in neurons. Voltage-operated, chemical-operated calcium channels and mechanisms of stability of intercellular calcium homeostasis (mitochondria, endoplasmic reticulum, calcium binding proteins, calcium exchange and calcium pump) are demonstrated in normal and pathological condition (105 ref.).     

Structural characteristics of the axons of nerve cells of the cat motor cortex

The axons of both pyramid and stellate neurons have a great number of ramifying collaterals which are considered in a comparative aspect. The both types of nerve cells are characterized by almost similar systems of the branching of axons, types of collaterals and end structures. The neurons are distinguished due to progressive development of one features of the axon structure and to a certain reduction of the others. In extreme forms the stellate cells have a well developed pericellular network, not coming outside the limits of the branching of their dendrites, while the pyramid cells are characterized by the mighty development of the main trunk and the absence of recurrent collaterals. Between these forms there are transitional forms.     

Anomalies of the branchial apparatus

Developmental abnormalities of the branchial apparatus represent a common cause of congenital lateral neck mass. Most often this is seen in the pediatric population but certainly may present at a much later age. The branchial apparatus contributes to major development of the head and neck, and a thorough understanding of embryology and anatomy is essential in treating these patients. Branchial anomalies may present as a cyst, sinus, or fistula; infection frequently complicates successful long-term management. Most often these lesions can be diagnosed by thorough history and physical examination. Complete surgical excision is the treatment of choice; however, recurrences may be encountered especially in those patients with a history of prior surgery or local infection.     

Posttraumatic detachment of the ciliary apparatus

Two new instruments are described for use in the cryotherapy of peripheral nerves. They are designed to give good vision and access, protect surrounding tissues and be comfortable to hold.