Vol.27-Issue 3- 2003
Ultrastructural effects of two phthalocyanines in CHO-K1 and
HeLa cells after laser irradiation
MARCELO DE CASTRO PAZOS*, CRISTINA PACHECO-SOARES*, NEWTON SOARES DA SILVA*,
RENATO AUGUSTO
DAMATTA**, AND MARCOS TADEU T. PACHECO*.
* Laboratório de Biologia Celular e Tecidual, Instituto de Pesquisa e Desenvolvimento, Universidade do Vale do Paraíba,
Av. Shishima Hifumi, 2911, CEP 12244-000, São José dos Campos, São Paulo, Brazil.
** Laboratório de Biologia Celular e Tecidual, Centro de Biociências e Biotecnologia, Universidade Estadual do Norte
Fluminense, Av. Alberto Lamego, 2000, CEP 28015-620, Campos dos Goytacazes, Rio de Janeiro, Brazil.
Key words: Photodynamic therapy, phthalocyanines, laser, cell death, ultrastructure.
ABSTRACT: The effects of Photodynamic Therapy using 2nd generation photosensitizers have been widely
investigated aiming clinical application treatment of solid neoplasms. In this work, ultrastructure changes
caused by the action of two 2nd generation photosensitizers and laser irradiation on CHO-K1 and HeLa (neoplastic)
cells were analyzed by transmission electron microscopy. Aluminum phthalocyanine chloride, aluminum
phthalocyanine tetrasulfonate chloride and radiation from a semiconductor laser at a fluency of 0.5 J/
cm2 (Power=26mW; l=670nm) were used. The results showed induction of apoptosis. Such alterations where
observed in HeLa but not in CHO-K1 cells after Aluminum phthalocyanine tetrasulfonate chloride (AlPcS4)
photodynamic treatment. The Aluminum phthalocyanine chloride (AlPc) photodynamic treatment induced
necrosis on the neoplastic cell line, and cytoplasm and nuclear alterations on the normal cell line.
Agrobacterium rhizogenes vs auxinic induction for in vitro
rhizogenesis of Prosopis chilensis and Nothofagus alpina
LUIS A. CARO1, NATALIA SANTECCHIA1, PABLO A. MARINANGELI1,
NÉSTOR R. CURVETTO1,2 AND LUIS F.
HERNÁNDEZ1,3
1. Departamento de Agronomía, Universidad Nacional del Sur, San Andrés 800, 8000 Bahía Blanca, Argentina.
2. CONICET (Consejo Nacional de Investigaciones Científicas y Técnicas), 1033 Buenos Aires, Argentina.
3. CIC (Comisión de Investigaciones Científ icas de la Provincia de Buenos Aires), 1900 La Plata, Argentina.
Key words: Agrobacterium rhizogenes, algarrobo, hairy roots, indole-3-butyric acid, Nothofagus nervosa, Prosopis chilensis, raulí, rooting
ABSTRACT: The induction and improvement of in vitro rhizogenesis of microshoots of Prosopis chilensis (Mol.) Stuntz and Nothofagus alpina (Poep. et Endl. Oerst.) were compared using Agrobacterium rhizogenes (Ar) versus indole-3-butyric acid (IBA) in the culture media. Microshoots of P. chilensis (1-2 cm length),
coming from in vitro grown seedlings, were cultivated in a modified Broadleaved Tree Medium (BTMm)
containing half salt concentration of macronutrients and 0.05 mg.L-1 benzilaminopurine (BAP). After 30
days, microshoots with 2-4 leaves were selected and cultured in BTMm-agar in presence or abscense of Ar
and in combination with IBA. For N. alpina, the apical shoots with the first 2 true leaves, from 5 weeks old
seedlings, were cultured in the abovementioned medium, but with 0.15 mgL-1 of BAP. After 2 months,
microshoots with 2-3 leaves were selected and cultured in BTMm-agar, supplemented with 5 mg.L-1 IBA or
in liquid BTMm on perlite and, in the presence or absence of A. rhizogenes (Ar) and in combination with 3
mg.L-1 IBA. Rooting in P. chilensis reached 100.0% when Ar infection was produced in the presence of IBA,
increasing both, the number and dry weight of roots. In N. alpina, 90.0% of rooting efficiency was obtained
when Ar infection was produced in liquid culture and in the absence of auxin.
Abbreviations: Ar, Agrobacterium rhizogenes; BAP, benzilaminopurine; BTMm, Broadleaved Tree Medium,
modified (half macronutrient, concentration); IBA, indole-3-butyric acid; MS, Murashige and Skoog medium;
PAR, photosynthecally active radiation.
Morphological study of the spermatogenesis in the teleost
Piaractus mesopotamicus
CARMINDA DA CRUZ-LANDIM*, FÁBIO CAMARGO ABDALLA* AND MARIA ALICE DA CRUZ-HÖFLING**
* Departamento de Biologia, Instituto de Biociências, Universidade Estadual Paulista (UNESP), Rio Claro, SP, Brazil.
** Departamento de Histologia e Embriologia, Instituto de Biologia, Universidade Estadual de Campinas (UNICAMP),
Campinas, SP, Brazil.
Keywords: Piaractus mesopotamicus, spermatogenesis, spermatogonia, spermiogenesis, Teleostei
ABSTRACT: The spermatogenesis of Piaractus mesopotamicus was investigated under light and transmission
electron microscopy. The specimens were captured from their natural environment (Rio Miranda and Rio
Aquidauana, Pantanal Matogrossense, Brazil) during April and September. The results were compared with
the spermatogenic data of specimens under captivity condition. In both conditions, P. mesopotamicus presented
the typical spermatogenesis pattern of the teleost f ishes, showing no significative differences. The
spermatozoon was classified as type I, which has a globular head without acrosome, a short middle piece and
a long tail constituted only by the flagellum. This type of spermatozoon is considered the basic type in fishes.
A unique mechanism of nuclear division in Giardia lamblia
involves components of the ventral disk and the nuclear envelope
ALBERTO J. SOLARI*, MONICA I. RAHN*, ALICIA SAURA**, AND HUGO D. LUJAN**
* Centro de Investigaciones en Reproduccion, Facultad de Medicina, UBA, Buenos Aires, Argentina.
** Catedra de Bioquímica y Biología Molecular, Facultad de Medicina, Universidad Nacional de Córdoba, Córdoba, Argentina
Key words: Giardia, primitive mitosis, fine structure of mitosis, protist mitosis, cell differentiation
ABSTRACT: The fine structure of the binucleate, parasitic protist Giardia lamblia during interphase and divisional stages was studied by serial thin sectioning and three-dimensional reconstructions. The earlier sign of nuclear division is the development of a few peripheral areas of densely packed chromatin directly attached to the inner nuclear envelope. An intracytoplasmic sheet of ventral disk components grows from the cell periphery towards one of the nuclei, apparently constricting this nucleus, which becomes located at a ventral bulge. After the basal bodies become duplicated, a full nuclear division occurs in trophozoites, giving two pairs of parent-daughter nuclei. This full division occurs in a dorsal-ventral direction, with the resulting nuclear pairs located at the sides of the two sets of basal bodies. A new ventral disk is formed from the diskderived sheets in the cell harboring the four nuclei. Cytokinesis is polymorphic, but at early stages is dorsalto-dorsal. Encysting trophozoites show the development of Golgi cisternae stacks and dense, specific secretory granules. 3-D reconstructions show that cysts contain a single pair of incompletely strangled nuclei. The dividing Giardia lacks a typical, microtubular spindle either inside or outside the nuclei. The nuclear envelope seems to be the only structure involved in the final division of the parent-daughter nuclei.
Goblet cell number in the ileum of rats denervated during
suckling and weaning
LUZMARINA HERNANDES*, LUCIENI CRISTINA MARQUES DA SILVA PEREIRA* AND ELIANA PARISI ALVARES**
* Departamento de Ciências Morfofisiológicas, Universidade Estadual de Maringá, Maringá, Paraná. Brazil.
** Departmento de Histologia e Embriologia, Instituto de Ciências Biomédicas I, Universidade de São Paulo, São Paulo,
São Paulo. Brazil.
Key words: benzalkonium chloride, development, goblet cells, myenteric plexus, small intestine.
ABSTRACT: The enteric nervous system plays a role on the stimulation of secretory cells of intestinal
epithelia. We have demonstrated that ablation of ENS stimulates epithelial cell proliferation. As goblet cells
are important constituents of the epithelial sheet, it is mandatory to investigate separately this cell type. The
myenteric plexus of the ileum of rats in postnatal development was partially removed by the serosal application
of benzalkonium chloride (BAC). Three groups of animals were used: those where BAC application was
at 13 days and sacrifice was at 15 (13/28-day-old) or 23 days after treatment (13/36-day-old), and those where
BAC was applied at 21 days and rats were killed 15 days after treatment (21/36-day-old) . The number of
goblet cells in the ileum was estimated in sections stained by periodic acid - Schiff (PAS) histochemistry. In
the 13/28 and 21/36 groups, the number of goblet cells was significantly higher after BAC treatment. These
results suggest that the myenteric denervation may have an acute effect on the number of goblet cell in
suckling and weanling rats, probably through submucous plexus.
Sperm ultrastructure and spermatogenesis in the lizard,
Tropidurus itambere
ADELINA FERREIRA AND HEIDI DOLDER
Departamento de Biologia Celular, Universidade Estadual de Campinas (Unicamp), Brazil
Keywords: ultrastructure, sperm, spermatogenesis, lizard, Tropiduridae
ABSTRACT: Spermatogenesis, with emphasis on spermiogenesis, is described for the lizard, Tropidurus
itambere, using light microscopy, phase contrast and epifluorescence, as well as scanning and transmission
electron microscopy. Cellular differentiation involves events of chromatin condensation, nuclear elongation
and the formation of structural complexes, such as the acrosomal and axonemal ones. Other new characteristics,
exclusive for this species, include various aspects of the subacrosomal granule, the insertion of the proacrosomal
vesicle and the development of these structures to participate in the acrosomal complex. Radial
projections occur just above the nuclear shoulders, which have been recognized already from the beginning of
cellular elongation. The development of the midpiece, the dense bodies, formation of the flagellum and elimination
of residual cytoplasm result in the final characterization of the mature spermatozoon. Comparisons
between Tropiduridae and other lizard families are made.
Edematogenic and myotoxic activities of the duvernoy’s gland
secretion of Philodryas olfersii from the north-east region of
Argentina
OFELIA ACOSTA DE PÉREZ1, LAURA LEIVA DE VILA2, MARÍA ELISA PEICHOTO2, SILVANA MARUÑAK3, RAQUEL
RUÍZ3, PAMELA TEIBLER3, CAROLINA GAY2 AND LAURA REY4
1. CONICET, Facultad de Ciencias Veterinarias, Universidad Nacional del Nordeste (UNNE). Sargento Cabral 2139, (3400)
Corrientes, Argentina.
2. Facultad de Ciencias Exactas y Naturales y Agrimensura, Universidad Nacional del Nordeste (UNNE). Corrientes, Argentina.
3. Facultad de Ciencias Veterinarias, Universidad Nacional del Nordeste (UNNE). Corrientes, Argentina.
4. Serpentario del Zoológico de la Ciudad de Corrientes. Corrientes, Argentina.
Keywords: Philodryas olfersii, edematogenic activity, myotoxic activity.
ABSTRACT: Philodryas olfersii is found in South America, from Amazonas to Patagonia. It is important to
characterize the venom of P. olfersii, who inhabits the North-East region of Argentina, since snake venoms
are known to exhibit considerable variability in composition and biological activities. In this work, mice
weighing 18-20 g (n = 4 for each experimental group) were used. For the edematogenic activity mice were
injected s.c. in the right foot pad with 50 ml of solutions containing different amounts of venom, whereas the
left foot pad was injected with 50 ml of PBS. Two hours after injection mice were killed by cervical dislocation
and both feet were cut off and weighed individually. For the myotoxic activity mice were injected i.m.
with 100 ml of solutions containing 40 mg of venom. Blood samples were extracted after 1, 3, 6, 8, 10, 12, 14,
16 and 24 h of venom injection to determinate serum CPK activity and mice were sacrif iced at the same time
intervals to obtain the inoculated gastrocnemius muscle. They were f ixed with Bouin solution and stained
with Hematoxylin-Eosin. Results showed that P. olfersii venom exhibits a high edematogenic activity (MED
= 0.31 mg) and a moderate myotoxic activity. Myonecrosis reached its highest level after 12 h of venom
injection as shown by plasmatic CPK levels (5,401 ± 330 IU/l) and microscopic assay. It demonstrates the
potential toxicity of the venom of P. olfersii, who inhabits the North-East region of Argentina. It also reinforces
the original warning concerning the potential danger of bites by colubrids.