mu(1)- and 5-HT1-dependent mechanisms in the anterior pretectal nucleus mediate the antinociceptive effects of retrosplenial cortex stimulation in rats

Aim: This study examines if injection of cobalt chloride (CoCl2) or antagonists of muscarinic cholinergic (atropine), mu(1)-opioid (naloxonazine) or 5-HT1 serotonergic (methiothepin) receptors into the dorsal or ventral portions of the anterior pretectal nucleus (APtN) alters the antinociceptive effects of stimulating the retrosplenial cortex (RSC) in rats. Main method: Changes in the nociceptive threshold were evaluated using the tail flick or incision pain tests in rats that were electrically stimulated at the RSC after the injection of saline, CoCl2 (1 mM, 0.10 mu L) or antagonists into the dorsal or ventral APtN. Key findings: The injection of CoCl2, naloxonazine (5 mu g/0.10 mu L) or methiothepin (3 mu g/0.10 mu L) into the dorsal APtN reduced the stimulation-produced antinociception from the RSC in the rat tail flick test. Reduction of incision pain was observed following stimulation of the RSC after the injection of the same substances into the ventral APtN. The injection of atropine (10 ng/0.10 mu L) or ketanserine (5 mu g/0.10 mu L) into the dorsal or ventral APtN was ineffective against the antinociception resulting from RSC stimulation. Significance: mu(1)-opioid- and 5-HT1-expressing neurons and cell processes in dorsal and ventral APtN are both implicated in the mediation of stimulation-produced antinociception from the RSC in the rat tail flick and incision pain tests, respectively. (c) 2012 Elsevier Inc. All rights reserved.

Biocompatibility and setting time of CPM-MTA and white Portland cement clinker with or without calcium sulfate

To evaluate the biocompatibility and the setting time of Portland cement clinker with or without 2% or 5% calcium sulfate and MTA-CPM. Twenty-four mice (Rattus norvegicus) received subcutaneously polyethylene tubes filled with Portland cement clinker with or without 2% or 5% calcium sulfate and MTA. After 15, 30 and 60 days of implantation, the animals were killed and specimens were prepared for microscopic analysis. For evaluation of the setting time, each material was analyzed using Gilmore needles weighing 113.5 g and 456.5 g, according to the ASTM specification Number C266-08 guideline. Data were analyzed by ANOVA and Tukey's test for setting time and Kruskal-Wallis and Dunn test for biocompatibility at 5% significance level. Histologic observation showed no statistically significant difference of biocompatibility (p>0.05) among the materials in the subcutaneous tissues. For the setting time, clinker without calcium sulfate showed the shortest initial and final setting times (6.18 s/21.48 s), followed by clinker with 2% calcium sulfate (9.22 s/25.33 s), clinker with 5% calcium sulfate (10.06 s/42.46 s) and MTA (15.01 s/42.46 s). All the tested materials showed biocompatibility and the calcium sulfate absence shortened the initial and final setting times of the white Portland cement clinker

Diethylpropion produces psychostimulant and reward effects

Diethylpropion (DEP) is a stimulant drug widely used for weight control in Brazil and other American countries. However, its effects on behavior and addiction potential are not yet well known. Data suggest that sensitization resulting from pre-exposure to psychostimulants could be a possible risk factor in subsequent drug addiction. The purpose of this investigation was to verify whether pre-exposure to DEP would sensitize rats to the motor activating effect and to the rewarding value of DEP. Two experiments were conducted. In both experiments rats were pre-exposed to DEP (20 mg/kg) or vehicle for 7 consecutive days. The acute effect of DEP (0.0, 1.0, 2.5 or 5.0 mg/kg) on motor activity (Experiment 1) and induction of Conditioned Place Preference-CPP (Experiment 2) were then measured. Results from Experiment 1 showed that 2.5 and 5.0 mg/kg DEP increased motor activity. Sensitization of this motor effect was observed. In Experiment 2, the doses of 2.5 and 5.0 mg/kg DEP induced CPP, indicating their rewarding value. However, no sensitization effect was observed. The results suggest that DEP at low doses has psychostimulant and rewarding properties. It is recommended that more effort should be dedicated to elucidating DEP abuse Potential. (c) 2008 Elsevier Inc. All rights reserved.

Epidural anesthesia and postoperatory analgesia with alpha-2 adrenergic agonists and lidocaine for ovariohysterectomy in bitches

The aim of this study was to determine the viability and cardiorespiratory effects of the association of epidural alpha-2 adrenergic agonists and lidocaine for ovariohysterectomy (OH) in bitches. Forty-two bitches were spayed under epidural anesthesia with 2.5 mg/kg body weight (BW) of 1% lidocaine with adrenaline (CON) or in association with 0.25 mg/kg BW of xylazine (XYL), 10 mu g/kg BW of romifidine (ROM), 30 mu g/kg BW of detomidine (DET), 2 mu g/kg BW of dexmedetomidine (DEX), or 5 mu g/kg BW of clonidine (CLO). Heart rate (HR), respiratory rate (f(R)) and arterial pressures were monitored immediately before and every 10 min after the epidural procedure. Blood gas and pH analysis were done before, and at 30 and 60 min after the epidural procedure. Animals were submitted to isoflurane anesthesia if they presented a slightest sign of discomfort during the procedure. Time of sensory epidural block and postoperative analgesia were evaluated. All animals in CON and DEX, 5 animals in ROM and CLO, 4 animals in XYL, and 3 in DET required supplementary isoflurane. All groups, except CLO, showed a decrease in HR. There was an increase in arterial pressures in all groups. Postoperative analgesia lasted the longest in XYL. None of the protocols were totally efficient to perform the complete procedure of OH; however, xylazine provided longer postoperative analgesia than the others.

Prevalence of Arterial Hypertension Among Seventh-Day Adventists of the Sao Paulo State Capital and Inner Area

Background: Inadequate life habits are known to favor hypertension, and Adventists recommend healthy life habits. Objective: To assess the prevalence of hypertension among Seventh-Day Adventists from the inner Sao Paulo state and Sao Paulo state capital. Methods: This study assessed 264 Adventists (mean age, 41.17 +/- 15.27 years; women, 59.8%) with a high religiosity level assessed by use of the Duke University Religion Index. Blood pressure was measured with a validated automatic device. The significance level adopted was p < 0.05. Results: The total prevalence of hypertension was 22.7% (27.4% in the inner state and 15% in the capital). The Adventists from the capital differed from those of inner state as follows (p < 0.05), respectively: higher education (62% vs 36.6%); employed by a third party (44%) vs self-employed (40.9%); family income (8.39 +/- 6.20 vs 4.59 +/- 4.75 minimum wages); individual income (4.54 +/- 5.34 vs 6.35 +/- 48; couple responsible for family income (35% vs 39.6%); vegetarianism (11% vs 3%); blood pressure (115.38 +/- 16.52/68.74 +/- 8.94 vs 123.66 +/- 19.62/74.88 +/- 11.85 mmHg); white ethnicity (65% vs 81.1%); married (53% vs 68.9%); lower tangible support in the social aspect (15.7 +/- 5.41 vs 16.9 +/- 4.32); and recalling the last time one's blood pressure was measured (65% vs 48.8%). On multivariate analysis, hypertension associated with the following: 1) vegetarianism (OR 0.051; 95% CI: 0.004-0.681); 2) educational level (OR 5.317; 95% CI: 1.674-16.893); 3) recalling the last time one's blood pressure was measured (OR 2.725; 95% CI: 1.275-5.821); 4) being retired (OR 8.846; 95% CI: 1.406-55.668); and 5) being responsible for family income (OR 0.422; 95% CI: 0.189-0.942). Conclusion: The prevalence of hypertension among Adventists was lower as compared with that reported in Brazilian studies, and it was lower in the Sao Paulo state capital as compared with that in the inner Sao Paulo state, possibly because of the better socioeconomic conditions and life habits of the former. (Arq Bras Cardiol 2012; 98(4): 329-337)

NLRP3 inflammasome-mediated neutrophil recruitment and hypernociception depend on leukotriene B4 in a murine model of gout

Objective Deposition of monosodium urate monohydrate (MSU) crystals in the joints promotes an intense inflammatory response and joint dysfunction. This study evaluated the role of the NLRP3 inflammasome and 5-lipoxygenase (5-LOX)derived leukotriene B4 (LTB4) in driving tissue inflammation and hypernociception in a murine model of gout. Methods. Gout was induced by injecting MSU crystals into the joints of mice. Wild-type mice and mice deficient in NLRP3, ASC, caspase 1, interleukin-1 beta (IL-1 beta), IL-1 receptor type I (IL-1RI), IL-18R, myeloid differentiation factor 88 (MyD88), or 5-LOX were used. Evaluations were performed to assess neutrophil influx, LTB4 activity, cytokine (IL-1 beta, CXCL1) production (by enzyme-linked immunosorbent assay), synovial microvasculature cell adhesion (by intravital microscopy), and hypernociception. Cleaved caspase 1 and production of reactive oxygen species (ROS) were analyzed in macrophages by Western blotting and fluorometric assay, respectively. Results. Injection of MSU crystals into the knee joints of mice induced neutrophil influx and neutrophildependent hypernociception. MSU crystal-induced neutrophil influx was CXCR2-dependent and relied on the induction of CXCL1 in an NLRP3/ASC/caspase 1/IL-1 beta/MyD88-dependent manner. LTB4 was produced rapidly after injection of MSU crystals, and this was necessary for caspase 1-dependent IL-1 beta production and consequent release of CXCR2-acting chemokines in vivo. In vitro, macrophages produced LTB4 after MSU crystal injection, and LTB4 was relevant in the MSU crystalinduced maturation of IL-1 beta. Mechanistically, LTB4 drove MSU crystal-induced production of ROS and ROS-dependent activation of the NLRP3 inflammasome. Conclusion. These results reveal the role of the NLRP3 inflammasome in mediating MSU crystalinduced inflammation and dysfunction of the joints, and highlight a previously unrecognized role of LTB4 in driving NLRP3 inflammasome activation in response to MSU crystals, both in vitro and in vivo.

The adipokine chemerin augments vascular reactivity to contractile stimuli via activation of the MEK-ERK1/2 pathway

Aims: Cytokines interfere with signaling pathways and mediators of vascular contraction. Endothelin-1 (ET-1) plays a major role on vascular dysfunction in conditions characterized by increased circulating levels of adipokines. In the present study we tested the hypothesis that the adipokine chemerin increases vascular contractile responses via activation of ET-1/ET-1 receptors-mediated pathways. Main methods: Male, 10-12 week-old Wistar rats were used. Endothelium-intact and endothelium-denuded aortic rings were incubated with chemerin (0.5 ng/mL or 5 ng/mL, for 1 or 24 h), and isometric contraction was recorded. Protein expression was determined by Western blotting. Key findings: Constrictor responses to phenylephrine (PE) and ET-1 were increased in vessels treated for 1 h with chemerin. Chemerin incubation for 24 h decreased PE contractile response whereas it increased the sensitivity to ET-1. Endothelium removal significantly potentiated chemerin effects on vascular contractile responses to PE and ET-1. Incubation with either an ERK1/2 inhibitor (PD98059) or ETA antagonist (BQ123) abolished chemerin effects on PE- and ET-1-induced vasoconstriction. Phosphorylation of MEK1/2 and ERK1/2 was significantly increased in vessels treated with chemerin for 1 and 24 h. Phosphorylation of these proteins was further increased in vessels incubated with ET-1 plus chemerin. ET-1 increased MEK1/2, ERK1/2 and MKP1 protein expression to values observed in vessels treated with chemerin. Significance: Chemerin increases contractile responses to PE and ET-1 via ERK1/2 activation. Our study contributes to a better understanding of the mechanisms by which the adipose tissue affects vascular function and, consequently, the vascular alterations present in obesity and related diseases. (c) 2012 Elsevier Inc. All rights reserved.

Leprosy reactions: coinfections as a possible risk factor

OBJECTIVE: This study aimed to determine the frequency of coinfections in leprosy patients and whether there is a relationship between the presence of coinfections and the development of leprosy reactional episodes. METHOD: A cross-sectional study based on an analysis of the medical records of the patients who were treated at the Leprosy Clinics of the Ribeirao Preto Medical School, University of Sao Paulo, was conducted from 2000 to 2010. Information was recorded regarding the age, sex, clinical status, WHO classification, treatment, presence of reactions and coinfections. Focal and systemic infections were diagnosed based on the history, physical examination, and laboratory tests. Multinomial logistic regression was used to evaluate the associations between the leprosy reactions and the patients' gender, age, WHO classification and coinfections. RESULTS: Two hundred twenty-five patients were studied. Most of these patients were males (155/225 = 68.8%) of an average age of 49.31 +/- 15.92 years, and the most prevalent clinical manifestation was the multibacillary (MB) form (n = 146), followed by the paucibacillary (PB) form (n = 79). Erythema nodosum leprosum (ENL) was more prevalent (78/122 = 63.9%) than the reversal reaction (RR) (44/122 = 36.1%), especially in the MB patients (OR 5.07; CI 2.86-8.99; p<0.0001) who exhibited coinfections (OR 2.26; CI 1.56-3.27; p<0.0001). Eighty-eight (88/225 = 39.1%) patients exhibited coinfections. Oral coinfections were the most prevalent (40/88 = 45.5%), followed by urinary tract infections (17/88 = 19.3%), sinusopathy (6/88 = 6.8%), hepatitis C (6/88 = 6.8%), and hepatitis B (6/88 = 6.8%). CONCLUSIONS: Coinfections may be involved in the development and maintenance of leprosy reactions.

The peripheral L-arginine-nitric oxide-cyclic GMP pathway and ATP-sensitive K+ channels are involved in the antinociceptive effect of crotalphine on neuropathic pain in rats

Crotalphine, a 14 amino acid peptide first isolated from the venom of the South American rattlesnake Crotalus durissus terrificus, induces a peripheral long-lasting and opioid receptor-mediated antinociceptive effect in a rat model of neuropathic pain induced by chronic constriction of the sciatic nerve. In the present study, we further characterized the molecular mechanisms involved in this effect, determining the type of opioid receptor responsible for this effect and the involvement of the nitric oxide-cyclic GMP pathway and of K+ channels. Crotalphine (0.2 or 5 mu g/kg, orally; 0.0006 mu g/paw), administered on day 14 after nerve constriction, inhibited mechanical hyperalgesia and low-threshold mechanical allodynia. The effect of the peptide was antagonized by intraplantar administration of naltrindole, an antagonist of delta-opioid receptors, and partially reversed by norbinaltorphimine, an antagonist of kappa-opioid receptors. The effect of crotalphine was also blocked by 7-nitroindazole, an inhibitor of the neuronal nitric oxide synthase; by 1H-(1,2,4) oxadiazolo[4,3-a]quinoxaline-1-one, an inhibitor of guanylate cyclase activation; and by glibenclamide, an ATP-sensitive K+ channel blocker. The results suggest that peripheral delta-opioid and kappa-opioid receptors, the nitric oxide-cyclic GMP pathway, and ATP-sensitive K+ channels are involved in the antinociceptive effect of crotalphine. The present data point to the therapeutic potential of this peptide for the treatment of chronic neuropathic pain. Behavioural Pharmacology 23:14-24 (C) 2012 Wolters Kluwer Health | Lippincott Williams & Wilkins.

Factors associated with preterm birth in Campina Grande, Paraiba State, Brazil: a case-control study

A case-control study (2008-2009) analyzed risk factors for preterm birth in the city of Campina Grande, Paraiba State, Brazil. A total of 341 preterm births and 424 controls were included. A multiple logistic regression model was used. Risk factors for preterm birth were: previous history of preterm birth (OR = 2.32; 95% CI: 1.25-4.29), maternal age (OR = 2.00; 95% CI: 1.00-4.03), inadequate prenatal care (OR = 2.15; 95% CI: 1.40-3.27), inadequate maternal weight gain (OR = 2.33; 95% CI: 1.45-3.75), maternal physical injury (OR = 2.10; 95% CI: 1.22-3.60), hypertension with eclampsia (OR = 17.08; 95% CI: 3.67-79.43) and without eclampsia (OR = 6.42; 95% CI: 3.50-11.76), hospitalization (OR = 5.64; 95% CI: 3.47-9.15), altered amniotic fluid volume (OR = 2.28; 95% CI: 1.32-3.95), vaginal bleeding (OR = 1.54; 95% CI: 1.01-2.34), and multiple gestation (OR = 22.65; 95% CI: 6.22-82.46). High and homogeneous prevalence of poverty and low maternal schooling among both cases and controls may have contributed to the fact that socioeconomic variables did not remain significantly associated with preterm birth.

Positive fluid balance is associated with reduced survival in critically ill patients with cancer

Background There are no studies that describe the impact of the cumulative fluid balance on the outcomes of cancer patients admitted to intensive care units ICUs. The aim of our study was to evaluate the relationship between fluid balance and clinical outcomes in these patients. Method One hundred twenty-two cancer patients were prospectively evaluated for survival during a 30-day period. Univariate (Chi-square, t-test, MannWhitney) and multiple logistic regression analyses were used to identify the admission parameters associated with mortality. Results The mean cumulative fluid balance was significantly higher in non-survivors than in survivors [1675?ml/24?h (4712921) vs. 887?ml/24?h (104557), P?=?0.017]. We used the area under the curve and the intersection of the sensibility and specificity curves to define a cumulative fluid balance value of 1100?ml/24?h. This value was used in the univariate model. In the multivariate model, the following variables were significantly associated with mortality in cancer patients: the Acute Physiology and Chronic Health Evaluation II score at admission [Odds ratio (OR) 1.15; 95% confidence interval (CI) (1.051.26), P?=?0.003], the Lung Injury Score at admission [OR 2.23; 95% CI (1.293.87), P?=?0.004] and a positive fluid balance higher than 1100?ml/24?h at ICU [OR 5.14; 95% CI (1.4518.24), P?=?0.011]. Conclusion A cumulative positive fluid balance higher than 1100?ml/24?h was independently associated with mortality in patients with cancer. These findings highlight the importance of improving the evaluation of these patients' volemic state and indicate that defined goals should be used to guide fluid therapy.

Quantitative Proteomic Analysis of the Effect of Fluoride on the Acquired Enamel Pellicle

The acquired enamel pellicle (AEP) is a thin film formed by the selective adsorption of salivary proteins onto the enamel surface of teeth. The AEP forms a critical interface between the mineral phase of teeth (hydroxyapatite) and the oral microbial biofilm. This biofilm is the key feature responsible for the development of dental caries. Fluoride on enamel surface is well known to reduce caries by reducing the solubility of enamel to acid. Information on the effects of fluoride on AEP formation is limited. This study aimed to investigate the effects of fluoride treatment on hydroxyapatite on the subsequent formation of AEP. In addition, this study pioneered the use of label-free quantitative proteomics to better understand the composition of AEP proteins. Hydroxyapatite discs were randomly divided in 4 groups (n = 10 per group). Each disc was exposed to distilled water (control) or sodium fluoride solution (1, 2 or 5%) for 2 hours. Discs were then washed and immersed in human saliva for an additional 2 hours. AEP from each disc was collected and subjected to liquid chromatography electrospray ionization mass spectrometry for protein identification, characterization and quantification. A total of 45 proteins were present in all four groups, 12 proteins were exclusively present in the control group and another 19 proteins were only present in the discs treated with 5% sodium fluoride. Relative proteomic quantification was carried out for the 45 proteins observed in all four groups. Notably, the concentration of important salivary proteins, such as statherin and histatin 1, decrease with increasing levels of fluoride. It suggests that these proteins are repulsed when hydroxyapatite surface is coated with fluoride. Our data demonstrated that treatment of hydroxyapatite with fluoride (at high concentration) qualitatively and quantitatively modulates AEP formation, effects which in turn will likely impact the formation of oral biofilms.

Cardiovascular effects of partial sleep deprivation in healthy volunteers

Dettoni JL, Consolim-Colombo FM, Drager LF, Rubira MC, de Souza SB, Irigoyen MC, Mostarda C, Borile S, Krieger EM, Moreno H Jr, Lorenzi-Filho G. Cardiovascular effects of partial sleep deprivation in healthy volunteers. J Appl Physiol 113: 232-236, 2012. First published April 26, 2012; doi: 10.1152/japplphysiol.01604.2011.-Sleep deprivation is common in Western societies and is associated with increased cardiovascular morbidity and mortality in epidemiological studies. However, the effects of partial sleep deprivation on the cardiovascular system are poorly understood. In the present study, we evaluated 13 healthy male volunteers (age: 31 +/- 2 yr) monitoring sleep diary and wrist actigraphy during their daily routine for 12 nights. The subjects were randomized and crossover to 5 nights of control sleep (>7 h) or 5 nights of partial sleep deprivation (<5 h), interposed by 2 nights of unrestricted sleep. At the end of control and partial sleep deprivation periods, heart rate variability (HRV), blood pressure variability (BPV), serum norepinephrine, and venous endothelial function (dorsal hand vein technique) were measured at rest in a supine position. The subjects slept 8.0 +/- 0.5 and 4.5 +/- 0.3 h during control and partial sleep deprivation periods, respectively (P < 0.01). Compared with control, sleep deprivation caused significant increase in sympathetic activity as evidenced by increase in percent low-frequency (50 +/- 15 vs. 59 +/- 8) and a decrease in percent high-frequency (50 +/- 10 vs. 41 +/- 8) components of HRV, increase in low-frequency band of BPV, and increase in serum norepinephrine (119 +/- 46 vs. 162 +/- 58 ng/ml), as well as a reduction in maximum endothelial dependent venodilatation (100 +/- 22 vs. 41 +/- 20%; P < 0.05 for all comparisons). In conclusion, 5 nights of partial sleep deprivation is sufficient to cause significant increase in sympathetic activity and venous endothelial dysfunction. These results may help to explain the association between short sleep and increased cardiovascular risk in epidemiological studies.

Fatores de risco pré-operatórios para mediastinite após cirurgia cardíaca: análise de 2768 pacientes

INTRODUÇÃO: A esternotomia mediana longitudinal é a via de acesso mais utilizada no tratamento das doenças cardíacas. As infecções profundas da ferida operatória no pós-operatório das cirurgias cardiovasculares são uma complicação séria, com alto custo durante o tratamento. Diferentes estudos têm encontrado fatores de risco para o desenvolvimento de mediastinite e as variáveis pré-operatórias têm tido especial destaque. OBJETIVO: O objetivo deste estudo é identificar fatores de risco pré-operatórios para o desenvolvimento de mediastinite em pacientes submetidos a revascularização do miocárdio e a substituição valvar. MÉTODOS: Este estudo observacional representa uma coorte de 2768 pacientes operados consecutivamente. O período considerado para análise foi de maio de 2007 a maio de 2009 e não houve critérios de exclusão. Foi realizada análise univariada e multivariada pelo modelo de regressão logística das 38 variáveis pré-operatórias eleitas. RESULTADOS: Nesta série, 35 (1,3%) pacientes evoluíram com mediastinite e 19 (0,7%) com osteomielite associada. A idade média dos pacientes foi de 59,9 ± 13,5 anos e o EuroSCORE de 4,5 ± 3,6. A mortalidade hospitalar foi de 42,8%. Na análise multivariada, foram identificadas três variáveis como preditoras independentes de mediastinite: balão intra-aórtico (OR 5,41, 95% IC [1,83 -16,01], P=0,002), hemodiálise (OR 4,87, 95% IC [1,41 - 16,86], P=0,012) e intervenção vascular extracardíaca (OR 4,39, 95% IC [1,64 - 11,76], P=0,003). CONCLUSÃO: O presente estudo demonstrou que necessidade do suporte hemodinâmico pré-operatório com balão intra-aórtico, hemodiálise e intervenção vascular extracardíaca são fatores de risco para o desenvolvimento de mediastinite após cirurgia cardíaca.

In vitro organogenesis of zucchini squash cv. Caserta

A protocol for the in vitro culture of Cucurbita pepo cv. Caserta was studied, using a cotyledon segment with an attached hypocotyl fragment as an explant. First, to determine the optimal seedling age, explants were collected from 4 to 6-day-old in vitro germinated seedlings and cultured in MS basal medium supplemented with benzylaminopurine (BAP, 4.5 mu M), under a 16-h photoperiod at 27 degrees C. Based on the results obtained, the explants collected from the 4-day-old seedlings were then cultured in MS basal medium supplemented with different concentrations of BAP (0, 1.1, 2.2, 3.3, 4.5, or 5.5 mu M) and incubated under a 16-h photoperiod at 27 degrees C. In vitro organogenesis was most efficient with explants collected from 4-day-old seedlings cultured in medium supplemented with 4.5 mu M of BAP. After 4 weeks of incubation the development of adventitious buds at the cotyledon/hypocotyl junction could be observed. These buds were transferred to elongation and rooting medium and the developed plants were acclimatized to greenhouse conditions. The morphogenic process was characterized using light and scanning electron microscopy analyses to confirm the organogenesis. The results showed that this alternate explant is efficient for in vitro culture of zucchini squash cv. Caserta. The protocol will be further examined for future use in genetic transformation experiments in this species.