Acetilación N-terminal, acetiltransferasas y mutaciones en subunidad auxiliar NAA15 de N-terminal acetiltransferasa A, asociadas a discapacidad intelectual y trastorno del espectro autista: una revisión bibliográfica / N-terminal acetylation, acetyltransferases and variants in NAA15 auxiliary subunit of the N-terminal acetyltransferase A associated with intellectual disability and autism spectrum disorder: a literature review

Las N-terminal acetiltransferasas (NaT) son fundamentales en el desarrollo, funcionamiento y vida media celular, acetilando gran parte del proteoma humano. Entre las ocho NaT identificadas, N-terminal acetiltransferasa A (NaTA) acetila a un mayor número de sustratos, teniendo además un rol fundamental en el neurodesarrollo. Previamente, estudios han demostrado que mutaciones en la subunidad catalítica de NaTA, NAA10, se asocian con trastornos del neurodesarrollo. Sin embargo, nuevas líneas investigativas sugieren que mutaciones de la subunidad auxiliar, NAA15, también tendrían un rol importante en el desarrollo de estos trastornos. Esta revisión se realiza con el objetivo de recopilar evidencia sobre variantes de NAA15 relacionadas con Discapacidad Intelectual (DI) y Trastorno de Espectro Autista (TEA). Se consultaron fuentes actualizadas sobre acetilación N-terminal, NaT, DI y TEA y mutaciones reportadas de NAA15 y sus expresiones fenotípicas, publicadas entre 2011 y 2022. Se concluye que, aun cuando existe relación entre mutaciones de NAA15, DI y TEA, todavía es necesario esclarecer los mecanismos fisiopatológicos de estos trastornos, el rol de NaTA y el impacto de variantes de sus subunidades en las vías moleculares y el fenotipo, lo que se dificulta por razones que van desde la complejidad de estas vías hasta el elevado costo de análisis genéticos. Se sugiere continuar la investigación en esta área, para comprender las bases moleculares subyacentes a estos trastornos y el rol de las mutaciones en subunidades de NaTA, con el fin último de estudiar potenciales tratamientos que mejoren la calidad de vida de las personas con estos trastornos y sus familias.

Nt-acetyltransferases (NaT) are essential in cell development, function and half-life, catalyzing most of the human proteome. Among the eight NaTs identified, N-terminal acetyltransferase A (NaTA) acetylates a greater number of substrates, also having a fundamental role in neurodevelopment. Previously, studies have shown that mutations in the catalytic subunit of NaTA, NAA10, are associated with neurodevelopmental disorders. However, new research lines suggest that mutations of the NAA15 helper subunit also plays an important role in the development of these disorders. This review is carried out with the objective of gathering evidence on NAA15 variants related to Intellectual Disability (ID) and Autism Spectrum Disorder (ASD). Updated sources on N-terminal acetylation, N-acetyltransferases, DI and TEA and reported mutations of NAA15 and their phenotypic expressions, published between 2011 and 2022 were consulted. It is concluded that even though there is a relationship between mutations of NAA15, ID and ASD exists, it is still necessary to clarify the pathophysiological mechanisms of these disorders, the role of NaTA and the impact of variants of its subunits in the molecular pathways and in the phenotype, for reasons ranging from the complexity of these pathways to the high cost of genetic testing. It is suggested to continue research in this area, to understand the molecular bases underlying these disorders and the role of mutations in NatA subunits, with the ultimate aim of studying potential treatments that improve the quality of life of people with these disorders and their families.

Preliminary study of ssDNA aptamer cytotoxicity on RhD+ red blood cells / 中国输血杂志

【Objective】 To investigate the effect of ssDNA aptamer of RhD blood group antigen on erythrocyte toxicity. 【Methods】 Two full-length ssDNA aptamers(82 bp) of RhD blood group antigen were obtained by gene synthesis.Five samples of whole blood with EDTA anticoagulant were collected to prepare red blood cell suspensions (4×107/mL), which were split into 10 tubes(100 μL/tube), corresponding to 5 experimental groups and 5 controls.Two monospecific full-length ssDNA sequences (100 pmol/μL, 5μL each) were added into the experimental group, while the same amount of normal saline into the control.After treatment, the experimental group and the control were incubated for 60 min at 37℃.After washing, they were suspended in LISS solution and stored at 4℃.The experimental group and the control were set according to different time point during storage (1 h, 1 d, 3 d, 10 d and 17 d), with 5 tubes in each group.For erythrocytes in LISS suspension at different storage time, Annexin V labeled with FITC was used as a probe to label the phosphatidylserine (PS) content and Fluo-4 to label Ca2+ .The eversion of PS and the change of Ca2+ concentration in red blood cells in LISS suspensions were determined by flow cytometry. 【Results】 After incubation, all groups were examined under the light microscope.No agglutination occurred in the experimental group, while agglutination occurred in the control.Flow cytometry showed that the number of Annexin V-FITC staining cells of suspended erythrocytes at the same storage time-point was similar between the experimental group and the control, with no significant differences.In the experimental group, apoptosis rate of Annexin V cells at 10-day storage(6.06±1.38) was significantly higher than that at 1-hour storage(P<0.05), so as at 17-day storage(7.77±1.23) than 1-hour, 1-day and 3-day storage(P<0.05). The apoptosis rate of Fluo-4 AM cell in suspended RBCs at the same storage time-point was similar between the two groups(P>0.05). In the experimental group, the apoptosis rate of Fluo-4 AM cell at the 3-day, 10-day and 17-day storage was 20.84±4.16, 22.35±3.37 and 27.06±2.81, respectively(P<0.05). 【Conclusion】 ssDNA aptamer was not found to have any cytotoxic effects on red blood cells, and RhD ssDNA aptamer may be used as a material for the detection and preparation of universal blood.

Improvement of germination rate and in vitro multiplication of Lavandula angustifolia

Lavender is well-known for its essential oils, which are in high demand and have a very important economicinterest, particularly for the pharmaceutical and cosmetic industries. This work had been done to study seedgermination and in vitro culture of Lavandula angustifolia. The seeds of L. angustifolia had undergonea vernalization followed by two treatments (physical and chemical), in order to increase the percentage ofgermination. Seeds were introduced into tubes containing the culture medium Murashige et Skoog (MS).Untreated seeds were germinated in the same medium supplemented with gibberellic acid at differentconcentrations. The kinetics, the speed, and the final germination rate were retained in order to evaluate theresponse of seeds. The micropropagation of L. angustifolia was performed on MS medium supplemented withdifferent concentrations of 6-Benzylaminopurine (Treatment I) and 6-Benzylaminopurine in combination withNaphthalene Acetic Acid (Treatment II). Multiplication rate, shoots number, nodes number, leaves number, andstem length were determined. Lavandula angustifolia seeds revealed very low germination percentages for allthe treatments used. However, those treated with sand (physical treatment) showed the highest percentage ofgermination (22%) followed by sulfuric acid treatment with a percentage of 14%. Results showed that the 11.11μM concentration of 6-Benzylaminopurine favored the maximum reactivity of the explants. The combinationbetween 6-Benzylaminopurine and Naphthalene Acetic Acid showed that MS supplemented with 8.88 μM6-Benzylaminopurine and 2.68 μM Naphthalene Acetic Acid was the most effective in the development ofvitroplants.

Altered thalamic neurotransmitters metabolism and functional connectivity during the development of chronic constriction injury induced neuropathic pain

BACKGROUND: To investigate the thalamic neurotransmitters and functional connections in the development of chronic constriction injury (CCI)-induced neuropathic pain. METHODS: The paw withdrawal threshold was measured by mechanical stimulation the right hind paw with the von frey hair in the rats of CCI-induced neuropathic pain. The N-acetylaspartate (NAA) and Glutamate (Glu) in thalamus were detected by magnetic resonance spectrum (MRS) process. The thalamic functional connectivity with other brain regions was scanned by functional magnetic resonance image (fMRI). RESULTS: The paw withdrawal threshold of the ipsilateral side showed a noticeable decline during the pathological process. Increased concentrations of Glu and decreased levels of NAA in the thalamus were significantly correlated with mechanical allodynia in the neuropathic pain states. The thalamic regional homogeneity (ReHo) decreased during the process of neuropathic pain. The functional connectivity among the thalamus with the insula and somatosensory cortex were significantly increased at different time points (7, 14, 21 days) after CCI surgery. CONCLUSION: Our study suggests that dynamic changes in thalamic NAA and Glu levels contribute to the thalamic functional connection hyper-excitation during CCI-induced neuropathic pain. Enhanced thalamus-insula functional connection might have a significant effect on the occurrence of neuropathic pain.

Micropropagation of Chapada Diamantina ornamental bromeliad / Micropropagação de bromélia ornamental da Chapada Diamantina

ABSTRACT: Sincoraea mucugensis (Wand. & A.A. Conc.) LOUZADA & WAND, an endangered bromeliad, is confined to the central region of the Chapada Diamantina, in the municipality of Mucugê, Brazil. From various researches, it is evident that for the propagation of this species, the in vitro technique is a feasible option. However, due to the low multiplication rates reported in various papers, this study aimed to establish a micropropagation protocol of direct organogenesis for S. mucugensis. First, the inoculation of the stem explants was done in MS ½ culture medium which contained different levels of BAP (0.00; 6.66; 8.88; 11.10; 13.20 µM) and NAA (0.00; 2.60; 5.20 µM). These shoots were then subjected to a couple of distinct rooting periods (of 30- and 60-day duration) using activated charcoal; finally, these microplants were transferred to a greenhouse for acclimatization, and covered with transparent plastic cups, as a water loss prevention test method. All the data were submitted to the analysis of variance (ANOVA), and the means were subjected to regression analysis or compared using the Tukey test. The findings revealed that the S. mucugensis stem explants raised in the NAA-rich medium (6.42 to 7.43 shoots/explants) showed high multiplication rates; the shoot rooting was done for 30 days using activated charcoal with the medium. Acclimatization, which was performed by directly exposing the microplants to the ex vitro environment, showed 95% survival rate.

RESUMO: Sincoraea mucugensis (Wand. & A.A. Conc.) LOUZADA & WAND é uma bromélia vulnerável de ocorrência restrita ao município de Mucugê, Chapada Diamantina. Estudos indicam que a cultura de tecidos é uma alternativa viável para a propagação in vitro desta espécie. Contudo, em função das baixas taxas de multiplicação obtidas em estudos anteriores, objetivou-se estabelecer um protocolo de micropropagação via organogênese direta para S. mucugensis. Explantes caulinares foram inoculados em meio de cultura MS ½ contendo diferentes concentrações de BAP (0,00; 6,66; 8,88; 11,10; 13,20 µM) e ANA (0,00; 2,60; 5,20 µM). Os brotos obtidos foram submetidos a diferentes períodos de enraizamento (30 e 60 dias) com carvão ativado; posteriormente as microplantas foram aclimatizadas em casa de vegetação, testando-se o efeito da cobertura com copos plásticos transparentes como estratégias contra perda de água. Os dados foram submetidos à análise de variância (ANOVA) e as médias analisadas por regressão ou comparadas pelo Teste de Tukey. Os resultados demonstram que altas taxas de multiplicação de S. mucugensis são geradas a partir de explantes caulinares cultivados em meio contendo ANA (6,42 a 7,43 brotos/explantes); o enraizamento dos brotos é realizado por 30 dias em meio com carvão ativado e a aclimatização é feita com exposição direta das microplantas ao ambiente ex vitro com 95% de sobrevivência.

Electroacupuncture ameliorate learning and memory by improving N -acetylaspartate and glutamate metabolism in APP/PS1 mice

OBJECTIVE: To explore the precise mechanism of electroacupuncture (EA) to delay cognitive impairment in Alzheimer disease. Methods N -Acetylaspartate (NAA), glutamate (Glu) and myoinositol (mI) metabolism were measured by magnetic resonance spectroscopy, learning and memory of APP/PS1 mouse was evaluated by the Morris water maze test and the step-down avoidance test, neuron survival number and neuronal structure in the hippocampus were observed by Nissl staining, and BDNF and phosphorylated TrkB detected by Western blot. RESULTS: EA at DU20 acupuncture significantly improve learning and memory in behavioral tests, up-regulate NAA, Glu and mI metabolism, increase the surviving neurons in hippocampus, and promote the expression of BDNF and TrkB in the APP/PS1 transgenic mice. CONCLUSION: These findings suggested that EA is a potential therapeutic for ameliorate cognitive dysfunction, and it might be due to EA could improve NAA and Glu metabolism by upregulation of BDNF in APP/PS1 mice.

Induction of in vitro shoots of Billbergia euphemiae E. Morren (Bromeliaceae) from leaf explants / Indução in vitro de brotos de Billbergia euphemiae E. Morren (Bromeliaceae) a partir de explantes foliares

Bromeliads are an important group for the maintenance of the Atlantic Forest, with many threatened species due to exacerbated extraction and destruction of their natural habitats. Considering the need of developing protocols for the conservation of these species, the aim of this work was to evaluate the effect of different growth regulators in the in vitro induction of shoots of Billbergia euphemiae. Leaf explants were excised from seedlings derived from in vitro germination and grown on MS medium supplemented with NAA (0, 1 or 2 µM) and BA (0, 2, 4 or 6 µ M) combinations. The evaluation of the number of shoots per explant, shoot length, number of leaves per shoot and longest leaf length average was carried out after 30 and 60 days of culture. The best in vitro responses were observed in the presence of 1 µM NAA after 60 days of culture, which induced the best production of shoots per explant (16.39), as well as the highest rates of shoot length (1.08 cm), number of leaves per shoot (5.00) and the longest leaf length (0.56 cm). This work determined the best conditions for shoot production from leaf explants of B. euphemiae, being the first report on micropropagation of this species.

As bromélias constituem um importante grupo para a manutenção da Floresta Atlântica, com várias espécies ameaçadas de extinção pelo extrativismo exacerbado e a destruição dos habitats naturais. Considerando a necessidade do desenvolvimento de protocolos para a conservação destas espécies, este trabalho teve como objetivo avaliar o efeito de diferentes reguladores de crescimento na indução de brotos de Billbergia euphemiae. Explantes foliares foram excisados de plântulas derivadas da germinação in vitro e inoculados em meio MS suplementado com combinações de ANA (0, 1 ou 2 µM) e BA (0, 2, 4 ou 6 µM). O número de brotos por explantes, comprimento dos brotos, número médio de folhas por broto e comprimento médio da maior folha foram avaliados após 30 e 60 dias de cultura. As melhores respostas foram observadas na presença de ANA a 1 µM, após 60 dias de cultura, que induziu a maior produção de brotos por explante (16,39), assim como as maiores taxas de comprimento dos brotos (1,08 cm), número médio de folhas por broto (5,00) e comprimento da maior folha (0,56 cm). Este trabalho determinou as melhores condições para produção de brotos de B. euphemiae a partir de explantes foliares, representando o primeiro relato de micropropagação desta espécie.

Micropropagation of Endangered and Decorative Species Dianthus pinifolius Sibth. et Sm

The aim of this study was to establish an efficient protocol for the in vitro propagation of the endangered, endemic and decorative species Dianthus pinifolius Sibth. et Sm. The effects of different concentrations of 6-benzylaminopurine (BAP) and naphtalenacetic acid (NAA), and different explant types (single-node cuttings, terminal buds and shoot cuttings) on shoot multiplication were examined on Murashige and Skoog (MS) and half-strength MS media. The best results were obtained for shoot cuttings on the MS medium supplemented with 0.5 mg/L BAP and 0.1 mg/L NAA, achieving a maximum rate of regeneration (100%) and a total of 15.4 newly-developed shoots per explant. The highest rooting rate (96.7%) was obtained on MS medium containing 1 mg/L NAA, while the acclimatization of the microplants obtained to ex vitro conditions was successful (88.9%).

The effect of Naa10p on the sensitivity of tongue squamous cell carcinoma Tca8113 cells to pingyangmycin / 实用口腔医学杂志

Objective:To study the effect of N-acetyltransferase 10(Naa10p)expression on the sensitivity of tongue squamous cell carcinoma Tca8113 cells to pingyangmycin(PYM).Methods:The lentiviral vectors of LV-shNaa10p,LV-Naa10p and control LV-NC were transfected into Tca8113 cells,the infection efficiency were identified,then the sensitivity of the infected cells to PYM was determined by MTS assay.Results:The IC50 of Tca8113-LV-shNaa10p group,Tca8113-LV-Naa10p group and Tca8113-LV-NC group were (20.772 ±0.106)μg/ml,(2.157 ±0.123)μg/ml and (6.301 ±0.069)μg/ml respectively(between groups P <0.05).Con-clusion:Knock down of Naa10p may reduce and over expression of Naa10p can increase the sensitivity of Tca8113 cells to PYM.

Correlation of Ki-67 expression with MRS metabolism in low-and high-grade cerebral gliomas / 中国肿瘤临床

objective:To investigate the relationship between magnetic resonance spectroscopy (MRS) metabolism and Ki-67 expres-sion in high-(HGG) and low-grade gliomas (LGG) by analyzing Ki-67 expression and HGG and LGG metabolites. Methods:We consid-ered 56 pathologically confirmed glioma cases in our hospital. The Ki-67 expression and the MRS metabolism parameters in the tu-mors were analyzed simultaneously. Results:The tumor solid value of Cho was positively correlated with the Ki-67 expression level (rs=0.714, P<0.05). By contrast, the Ki-67 expression level was negatively correlated with the tumor solid value of NAA (rs=?0.708, P<0.05) in 35 cases of the LGG group. The tumor solid value of Cho was also positively correlated with the Ki-67 expression level (rs=0.624, P<0.05). By comparison, the Ki-67 expression level was negatively correlated with the tumor solid value of NAA in the HGG group (rs=?0.769, P<0.05). Conclusion:The MRS metabolism was correlated with the Ki-67 expression in high-and low-grade gliomas.

In Vitro Propagation of Chamomilla Recutita From Capitulum Inflorescence: A Medicinal Plant With Multiple Therapeutic Applications

A protocol has been developed for induction of somatic embryogenesis from whole inflorescence explants of Chamomilla recutita L. (chamomile). Chamomile is a well-known medicinal plant from the Asteraceae family often referred to as the “star among medicinal species.” Nowadays, it is a highly favoured medicinal plant in folk and traditional medicine. Its multitherapeutic, cosmetic and nutritional values have been established through the years of traditional and scientific use and research. Chamomile has an established domestic (Indian) and international market, which is increasing day by day. Among the various major constituents, α-bisabolol and chamazulene have been reported to be more useful than others. Chamazulene occurs in the capitula of the flowers in minute quantities and has been demonstrated to exert antiinflammatory activity in-vivo. Moreover, chamomile is a seasonal 4-5 months winter crop in India but is extensively required in various medicinal applications. Therefore, to increase the overall yield of this plant, its in-vitro propagation is needed. In the present study, somatic embryos were developed from capitulum explants after 2-4 weeks of culture on MS medium supplemented with 26.8 µM NAA and 11.5 µM Kin. The somatic embryos were further subcultured in-vitro, where new plantlets regenerated from embryos. It is concluded that in-vitro propagation is possible in case of chamomile and can be used to increase the overall yield of chamazulene present in the capitula of flowers as well as augment the overall yield of this important plant, which is conventionally propagated by seeds.

Response of Pigeon Pea Landraces [Cajanus cajan (L.) Millsp.] to Exogenous Application of Plant Growth Regulators.

Aim: Optimizing plant growth regulators in the manipulation of growth physiology, development and seed yield of pigeon pea landraces is cardinal, especially as it complements other improvement techniques. The research was aimed at evaluating the effect of IAA, NAA and paclobutrazol singly and in combination, but especially paclobutrazol in reducing plant height. Methods: Thirty (30) seeds each of brown “Fiofio” [Cajanus cajan (L.) Millsp.] were soaked in 0, 100, and 150mg/l concentration of IAA, NAA, paclobutrazol, paclobutrazol + IAA and paclobutrazol + NAA, respectively for 48 hours. Results: Results obtained revealed that treating pigeon pea seeds with paclobutrazol caused reduction in plant height and inter-node length, which did not translate to higher yield. However, plants raised from pigeon pea seeds soaked in 100 and 150 mg/l paclobutrazol + NAA did excellently well in both yield and yield – related traits. Conclusion: The implication of these findings is that though treating seeds with paclobutrazol caused significant reduction in plant height and increased branch numbers, it needed to interact with NAA to cause holistic improvement, especially in seed yield.

In Vitro Regeneration from Shoot Tip and Nodal Explants of Simarouba Glauca DC, a Promising Biodiesel Tree.

An efficient regeneration protocol was developed from shoot tip and nodal explants of Simarouba glauca DC, a promising biodiesel plant. Nodal explants appeared to have better regeneration capacity than shoot tip explants (40%) in the tested media. The highest regeneration frequency (90%) and shoot number (7.00 ± 1.00 shoots per explants) were obtained in nodal explants in Murashige and Skoog’s (MS) medium supplemented with 6-benzylaminopurine (BAP) 4.43 μM and α-naphthalene acetic acid (NAA) 5.36 μM.Induced shoot buds were multiplied and elongated on the MS medium supplemented with BAP (4.44 μM), NAA (5.36 μM) and TDZ (Thidiazuron) 2.27 μM with 9.66±0.33 (mean length 5.35±0.32 cm) and 9.00±0.57 (mean length 4.51±0.15cm) shoots using nodal segments and shoot tip explants, respectively. Halfstrength woody plant medium (WPM) containing 2.46μM indole-3-butyric acid (IBA) produced the maximum number of roots (6.00±1.15). The rooted plantlets were hardened on MS basal liquid medium and subsequently in polycups containing sterile soil and vermiculite (1:1) and successfully established in pots.

Aplicação de extratos de folhas e tubérbulos de Cyperus rotundus L. e de auxinas sintéticas na estaquia caulinar de Duranta repens L. / Effect of Cyperus rotundus L. leaf and tuber extracts and synthetic auxins applied to stem cuttings of Duranta repens L.

É de conhecimento popular que extratos de tiririca aumentam o enraizamento adventício de várias espécies, sendo esse procedimento utilizado na propagação vegetativa caseira. Uma vez que nesses extratos concentram grandes quantidades de auxinas e compostos fenólicos, o objetivo deste estudo pioneiro foi avaliar o efeito da aplicação de extratos de folhas e de tubérculos de Cyperus rotundus L. na estaquia caulinar de Duranta repens L., espécie considerada de fácil enraizamento, comparando sua ação à de auxinas sintéticas. As estacas foram coletadas em abril, junho, agosto, outubro e dezembro/2006 e fevereiro/2007 e confeccionadas com 8 cm de comprimento, com base cortada em bisel e corte reto acima da última gema, mantendo-se 4 folhas apicais. Os tratamentos foram: água e solução alcoólica 50% como controles; extrato de folhas e de tubérculos de C. rotundus (25%, 50% e 100%); solução de ANA e AIB (500 e 1000 mg L-1), com imersão da base das estacas durante 10 segundos. As estacas foram mantidas sob nebulização, em tubetes contendo vermiculita, por 45 dias. Foram avaliadas a porcentagem de estacas enraizadas, número de raízes por estaca, comprimento das três maiores raízes por estaca (cm) e a porcentagem de estacas vivas. Estacas coletadas nos meses de outubro e dezembro/2006 apresentaram as maiores porcentagens de enraizamento (87,3% e 86,7%, respectivamente) e maior número de raízes por estacas (9,5 e 10,4 raízes, respectivamente). O comprimento máximo das 3 maiores raízes (10,2 cm) foi obtido em estacas coletadas em dezembro/2006 e a maior taxa de sobrevivência (60%) foi registrada na coleta de junho/2006 em estacas tratadas com 500 e 1000 mg L-1 de AIB. A aplicação dos extratos de folhas e de tubérculos de C. rotundus não apresentou diferença entre os resultados obtidos com a aplicação de ANA e AIB, os quais, por sua vez, também não influenciaram o enraizamento de estacas de D. repens em nenhuma das épocas avaliadas.

It is known from popular tradition that extracts from nut grass increase the adventitious rooting of different species, being this procedure used in home-made vegetative propagation. Since in these extracts it is possible to find a large amount of auxins and phenolic compounds, the objective of this study was to evaluate the effect of the application of Cypreus rotundus L. leaves and tubers extracts in stem cuttings of Duranta repens L., a species considered of easy rooting, in comparison with the action of synthetic auxins. Cuttings were collected in april, june, august, october and december/2006 and february/2007 with 8 cm in length, diagonal cut at the bottom and a straight cut above the last bud, keeping four upper leaves. Treatments consisted of control (water and ethanol 50%); extract from C. rotundus leaves and tubers (25, 50 and 100%); NAA and IBA (500 and 1000 mg L-1), with immersion of the base of the cuttings for 10 seconds. Cuttings were kept in greenhouse under high moisture, in plastic containers containing vermiculite, for 45 days. The percentage of rooted cuttings, number of roots per cutting, length of the three longest roots per cutting (cm) and the percentage of living cuttings were evaluated. Cuttings collected in october and december/2006 demonstrated the highest percentages of rooting (87.3% and 86.7%, respectively) and highest number of roots per cutting (9.5 and 10.4 roots, respectively). December/2006 cuttings showed the maximum length of the three largest roots (10.2 cm) and the best level of survival (60%) was found in cuttings collected in june/2006 and treated with 500 and 1000 mg L-1 of IBA. Therefore, we observed no statistical differences between the application of C. rotundus leaf and tuber extracts and the application of NAA and IBA.

In vitro Regeneration of Plantlets from Leaf and Nodal explants of Aristolochia indica L.- An Important Threatened Medicinal Plant

Objective: An efficient reproducible protocol has been developed for in vitro regeneration of plantlets from leaf and nodal explants of Aristolochia indica L. Methods: Wild grown plants Aristolochia indica L. were collected and grown in the departmental garden. Leaf and nodal segments (0.5-1.0 cm) from young healthy plants were first washed thoroughly under running tap water for 15 - 20 minutes and then treated with liquid detergent [5% (v/v) Tween-20] for 5-10 minutes. Later these explants were washed with double-distilled water for 5 minutes. Subsequently, explants were immersed in 70% (v/v) ethanol for 2 - 3 minutes and washed with sterile glass double distilled water for 2-3 times. Eventually, the explants were treated with an aqueous solution of 0.1% (w/v) HgCl2 for 1 - 2 minutes and rinsed for two-to-three times in sterile ddH2O to remove all traces of HgCl2. The sterilized explants were inoculated aseptically onto solid basal Murashige and Skoog’s medium with different concentrations and combinations of BAP and NAA for in vitro regeneration of plants. Results: Both leaf and nodal explants cultured on MS medium supplemented with 0.8 mg/L BAP developed into mass of callus. These calli were subcultured for the induction of shoots and roots. Shoots were induced from both calli on MS medium supplemented with 0.8 mg/L BAP+0.5 mg/L NAA. Roots were induced from in vitro shoots on MS medium supplemented with 0.8 mg/L NAA for 4 weeks. Nodal explants were more regenerative with 95 % response compared to leaf explants with 85%. Finally, these in vitro regenerated plantlets were hardened, acclimatised and successfully transferred to the field. Conclusions: The present protocol for in vitro regeneration of Aristolochia indica L. can be used to make this plant available throughout the year for traditional healers, pharmaceutical usages, germplasm conservation, commercial cultivation, and also for the production of secondary metabolites.

In Vitro Regeneration of Exacum wightianum Arn. (Gentianaceae)- An Endemic Medicinal Plant From The Nilgiris, Western Ghats.

Exacum wightianum Arn. (Gentianaceae) is an endemic medicinal plant from the Nilgiri hills, Western Ghats, Tamil Nadu. Indirect regeneration of E. wightianum was obtained through organogenesis in callus culture. Axial bud explants were found to be best suited for callus induction on MS medium supplemented with BA + NAA (2.0+0.03 mg/L). Multiple shoots originated from callus obtained from the axial buds. were multiplied by subculture on the same medium. Maximum shoot regeneration was obtained on MS medium supplemented with BA with NAA (2.0+0.5 mg/L), and up to 25shoots was observed within 2 weeks.

Micropropagation and shoot regeneration of Cistus creticus ssp. Creticus.

Cistus creticus L. ssp. creticus is a medicinal aromatic shrub native in Crete (Greece). The protocol described in this paper provides optimal levels of growth regulators required to obtain high regeneration rates of Cistus in vitro. Micropropagation has been achieved through rapid proliferation of shoot-tips on Murashige and Skoog (MS) basal medium supplemented with 0.2 mg l-1 6-benzylaminopurine (BAP). After four weeks shoots were transferred to MS medium without growth regulators for further development and rooting. The highest percentage of regenerated shoots was obtained with 0.1 mg l-1 TDZ and 0.1 mg l-1 NAA after 4 weeks. Elongation and rooting was readily achieved when multiple shoots more than 1 cm in length were singled out and cultured on the MS medium without growth regulators. The plantlets were successfully adapted and grew vigorously in greenhouse conditions. This is the first report of shoot regeneration in the genus Cistus. The regeneration protocol developed in this study provides a basis for further investigation of the medicinally active constituents of this elite medicinal plant.

Effects of growth regulator and culture methods on rooting of Ajuga lobata and content of β-ecdysone / 中草药

Objective: To study the effects of indole butyric acid (IBA) and NAA at different concentrations and culture methods on the growth of Ajuga lobata root and content of β-ecdysone. Methods: Taking MS as basic medium supplemented with IBA (0.5, 1.0, and 2.0 mg/L) or NAA (0.3, 0.6, and 0.9 mg/L), the tissue culture was carried out. HPLC was used to determinate the content of β-ecdysterone. Results: The results showed that the roots could grew well on MS as basic medium supplemented with IBA 0.5-1 mg/L. The best culture condition for the highest β-ecdysone accumulation was MS medium supplemented with IBA 2 mg/L. The content of β-ecdysterone in the whole plant and the roots of A. lobata with tissue culture was twice times of hydroponics and about three times of cultivation. Conclusion: MS medium supplemented with IBA at the proper concentration could promote the root growth of A. lobata. The content of β-ecdysterone in the root of A. lobata is the highest in tissue cultures.

Proton magnetic resonance spectroscopy of brain in obstructive sleep apnoea in north Indian Asian subjects.

Background & objectives: Repeated apnoeic/hypoapnoeic episodes during sleep may produce cerebral damage in patients with obstructive sleep apnoea (OSA). The aim of this study was to determine the absolute concentration of cerebral metabolites in apnoeic and non-apnoeic subjects from different regions of the brain to monitor the regional variation of cerebral metabolites. Methods: Absolute concentration of cerebral metabolites was determined by using early morning proton magnetic resonance spectroscopy (1H MRS) in 18 apnoeic patients with OSA (apnoeics) having apnoea/hypopnoea index (AHI) >5/h, while 32 were non-apnoeic subjects with AHI< 5/h. Results: The absolute concentration of tNAA [(N-acetylaspartate (NAA)+N-acetylaspartylglutamate (NAAG)] was observed to be statistically significantly lower (P<0.05) in apnoeics in the left temporal and left frontal gray regions compared to non-apnoeics. The Glx (glutamine, Gln + glutamate, Glu) resonance showed higher concentration (but not statistically significant) in the left temporal and left frontal regions of the brain in apnoeics compared to non-apnoeics. The absolute concentration of myo-inositol (mI) was significantly high (P<0.03) in apnoeics in the occipital region compared to non-apnoeics. Interpretation & conclusions: Reduction in the absolute concentration of tNAA in apnoeics is suggestive of neuronal damage, probably caused by repeated apnoeic episodes in these patients. NAA showed negative correlation with AHI in the left frontal region, while Cho and mI were positively correlated in the occipital region and Glx showed positive correlation in the left temporal region of the brain. Overall, our results demonstrate that the variation in metabolites concentrations is not uniform across various regions of the brain studied in patients with OSA. Further studies with a large cohort of patients to substantiate these observations are required.

Cloreto de sódio e ácido naftalenoacético no enraizamento de microestacas de amoreira-preta cv. Brazos in vitro / Sodium chloride and naphthaleneacetic acid on in vitro rooting of blackberry cv. Brazos

A micropropagação de amoreira-preta (Rubus sp.) é utilizada, principalmente, para a obtenção de plantas livres de vírus e num curto espaço de tempo. No presente trabalho foram testadas diferentes concentrações de cloreto de sódio (NaCl) e do ácido naftalenoacético (ANA)adicionados ao meio de cultura in vitro de amoreira-preta. O meio foi constituído de sais MS, acrescidos de 30 g L-1 de sacarose e 6 g L-1 de ágar, e o pH ajustado para 5,8 antes da autoclavagem a 121ºC e 1 atm por 20 minutos. Os tratamentos consistiram em concentrações de NaCl (0; 25; 50; 75 e 100 mg L-1) e de ANA (0; 0,1; 0,5; 1,0 e 1,5 mg L-1), em todas as combinações possíveis e da amoreira-preta cv. Brazos. Segmentos nodais, oriundos de plântulas preestabelecidas in vitro foram excisados e introduzidos em tubos de ensaio contendo 15 mL do meio de cultura. Posteriormente, os tubos foram transferidos para sala de crescimento a 25 ± 2ºC, irradiância de 35 mmol.m-2.s-1 e fotoperíodo de 16 horas. O delineamento experimental utilizado foi inteiramente casualisado, utilizando-se 4 repetições com 12 plântulas cada. O experimento foi avaliado após 70 dias de cultivo in vitro. O desenvolvimento in vitro da cv. Brazos foi favorecido nos tratamentos com 50-75 mg L-1 de NaCl e 1,0-1,5 mg L-1 de ANA, enquanto melhor resultado de enraizamento foi obtido no tratamento com 100 mg L-1 de NaCl e 1,5 mg L-1 de ANA.

The blackberry (Rubus sp.) micropropagation is used, mainly for obtaining virus-free plants in short period of time. In the present work different concentrations of sodium chloride (NaCl) and naphthaleneacetic acid (NAA) were tested added to in vitro culture medium of blackberry cv. Brazos. The culture medium consisted of MS salts, with 30 g L-1 sucrose 6 g L-1 agar and pH adjusted to 5.8 before the sterilization at 121ºC and 1 atm for 20 minutes. The treatments consisted of NaCl (0; 25; 50; 75 and 100 mg L-1) and NAA (0; 0.01; 0.5; 1.0 and 1.5 mg L-1) concentrations, in all possible combinations. Nodal segments, originating from plants established in vitro were excised and introduced in tubes containing 15 mL of culture medium. After that, the tubes were transferred to growth room at 25 ± 2ºC, irradiance of 35 mmol m-2 s-1 and photoperiod 16 hours. A completely randomizel block design with four replicates and 12 plants per replicate was used. The experiment was evaluated after 70 days of in vitro cultivation. The in vitro development of the cv. Brazos was favored in the treatments with 50 to 75 mg L-1 NaCl and 1.0 to 1.5 mg L-1 NAA, while the best rooting results were obtained using 100 mg L-1 NaCl and 1.5 mg L-1 NAA.