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1.
Hong Kong Med J ; 29(6): 489-497, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-38111367

RESUMO

INTRODUCTION: Healthcare workers in intensive care units often experience moral distress, depression, and stress-related symptoms. These conditions can lower staff retention and influence the quality of patient care. This study aimed to evaluate the prevalence of moral distress and psychological status among healthcare workers in a newly established paediatric intensive care unit (PICU) in Hong Kong. METHODS: A cross-sectional questionnaire survey was conducted in the PICU of the Hong Kong Children's Hospital; healthcare workers (doctors, nurses and allied health professionals) were invited to participate. The Revised Moral Distress Scale (MDS-R) Paediatric Version and Depression Anxiety and Stress Scale-21 items were used to assess moral distress and psychological status, respectively. Demographic characteristics were examined in relation to moral distress, depression, anxiety, and stress scores to identify risk factors for poor psychological outcomes. Correlations of moral distress with depression, anxiety, and stress were examined. RESULTS: Forty-six healthcare workers completed the survey. The overall median MDS-R moral distress score was 71. Nurses had a significantly higher median moral distress score, compared with doctors and allied health professionals (102 vs 47 vs 20). Nurses also had the highest median anxiety and stress scores (11 and 20, respectively). Moral distress scores were correlated with depression (r=0.445; P=0.002) and anxiety scores (r=0.417; P<0.05). Healthcare workers intending to quit their jobs had significantly higher moral distress scores (P<0.05). CONCLUSION: Among PICU healthcare workers, nurses had the highest level of moral distress. Moral distress was associated with greater depression, anxiety, and intention to quit. Healthcare workers need support and a sustainable working environment to cope with moral distress.


Assuntos
Pessoal de Saúde , Unidades de Terapia Intensiva Pediátrica , Humanos , Criança , Estudos Transversais , Unidades de Terapia Intensiva , Assistência ao Paciente , Inquéritos e Questionários , Princípios Morais , Estresse Psicológico/epidemiologia , Estresse Psicológico/etiologia
2.
Conscious Cogn ; 111: 103508, 2023 05.
Artigo em Inglês | MEDLINE | ID: mdl-37004356

RESUMO

Until now, most studies investigating the relationship between event segmentation and memory have used videos filmed from a third-person perspective, although people experience their lives from a first-person perspective. The present study aimed to determine whether visual perspective impacts events segmentation and further recall. Fifty-seven participants were recruited and assigned to either first- (1PP) or third-person perspective (3PP) condition, before segmenting videos of daily life activities. Our results showed that the although the number of event boundaries was higher in the 3PP condition than in the 1PP, no differences were observed for event segmentation qualitative abilities and organization. Memory of temporal order was better for events encoded in the 3PP than in the 1PP, while memory content was similar in both conditions. Higher event segmentation rates were correlated with a better recall of small actions and temporal order.


Assuntos
Rememoração Mental , Humanos , Estimulação Luminosa/métodos , Análise de Variância
3.
Acta Anaesthesiol Scand ; 58(6): 743-50, 2014 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-24734787

RESUMO

BACKGROUND: We studied the hypothesis that an equal spinal anaesthetic dose administered in the sitting position to patients undergoing post-partum tubal ligation (PPTL) and caesarean section (CS) would yield similar sensory block characteristics and analgesic efficacy. METHODS: This prospective, non-randomised trial recruited 20 women undergoing PPTL within 48 h of vaginal delivery and 20 undergoing CS. Spinal anaesthesia comprising intrathecal hyperbaric bupivacaine 12 mg and morphine 100 µg was administered at L3/4 with patients sitting. Our primary end point was the maximal dermatomal sensory block (to cold). RESULTS: Baseline demographics were comparable, but PPTL patients had greater parity, with mean ± standard deviation 17.54 ± 11.2 h from delivery to spinal anaesthesia, and shorter duration of surgery, 17.54 ± 11.2 vs. 40.3 ± 15.5 min. Similar maximal sensory blocks (to cold) were achieved in group PPTL vs. CS, T4 (T1-T5) vs. T3 (T1-T5), P = 0.104, in comparable times, 8.6 ± 2.6 vs. 7.6 ± 3.0 min, P = 0.267. PPTL patients had significantly faster two-segment block regression (70.7 ± 23.5 vs. 97.6 ± 23.9 min, P = 0.001) and to T10 (120.8 ± 35.6 vs. 145.1 ± 24.3 min, P = 0.016), with less hypotension (25% vs. 65%, P = 0.025) and phenylephrine (20.0 ± 60.6 µg vs. 120.0 ± 119.6 µg, P = 0.005). CONCLUSION: The same dose of hyperbaric bupivacaine 12 mg and morphine 100 µg administered in the sitting position to both PPTL and CS parturients yielded similar maximal sensory blocks, but PPTL exhibited faster block regression and less hypotension/vasopressor requirement.


Assuntos
Anestesia Obstétrica/métodos , Raquianestesia/métodos , Anestésicos Locais/administração & dosagem , Bupivacaína/administração & dosagem , Cesárea , Posicionamento do Paciente , Esterilização Tubária , Adulto , Período de Recuperação da Anestesia , Temperatura Baixa , Parto Obstétrico , Feminino , Humanos , Hipotensão/induzido quimicamente , Complicações Intraoperatórias/induzido quimicamente , Laparotomia , Morfina/administração & dosagem , Entorpecentes/administração & dosagem , Duração da Cirurgia , Período Pós-Parto , Gravidez , Estudos Prospectivos
4.
Asian J Surg ; 32(3): 143-50, 2009 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-19656753

RESUMO

OBJECTIVE: Patient satisfaction is an important indicator of healthcare system performance. High patient satisfaction is associated with greater trust in caregivers, improved compliance with treatment recommendations and a better quality of life (QOL). There are few validated instruments to measure surgical patients satisfaction. The aim of this study was to develop a culturally-specific patient satisfaction instrument, for use as an outcome measure in evaluating surgical services. DESIGN: Patient focus groups were convened to explore dimensions of the peri operative hospital experience. Forums uncovered pertinent domains of interest and identified terminology understood by patients. A preliminary set of items reflecting patient satisfaction was developed. Test-retest reliability of a new surgical patient satisfaction instrument was assessed in 42 subjects at hospital discharge. RESULTS: Domains that emerged included; admission processes and hospital environment, information provision, nursing care, doctor and nurse interaction, and ancillary staff services. Staff attitudes and human qualities were highly valued, as was prompt attention to requests for assistance. Clarity or quality of medical information did not appear to influence in-patient satisfaction. A new measure of surgical patient satisfaction, Hong Kong Index of Inpatient Happiness (HK2Happ), was developed from focus group consultation. Test-retest generated an Intra Class Correlation of 0.868-0.935, indicating a highly stable tool. CONCLUSION: The initial version of HK2Happ was reliable in assessing surgical patient satisfaction. The measure is now undergoing validity testing across different surgical patient populations for generalization and generation of a short form of discriminant items.


Assuntos
Cirurgia Geral/normas , Satisfação do Paciente , Assistência Perioperatória/normas , Qualidade da Assistência à Saúde , Inquéritos e Questionários , Povo Asiático , Características Culturais , Feminino , Grupos Focais , Hong Kong , Hospitalização , Humanos , Pacientes Internados , Masculino , Pessoa de Meia-Idade , Avaliação de Resultados em Cuidados de Saúde , Assistência ao Paciente/normas , Psicometria , Reprodutibilidade dos Testes , Resultado do Tratamento
5.
Hong Kong Med J ; 14(3): 192-7, 2008 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-18525087

RESUMO

OBJECTIVE: To summarise our experience of laparoscopic radical prostatectomy in a single centre in Hong Kong over 5 years. DESIGN: Retrospective study. SETTING: Urology Division, Department of Surgery, Tuen Mun Hospital, Hong Kong. PATIENTS: A total of 87 patients who underwent laparoscopic radical prostatectomy from March 2002 to May 2007. MAIN OUTCOME MEASURES: Peri-operative data and follow-up information. RESULTS: The operative procedure used entailed Montsouris technique and its modifications, including the latest method involving the extraperitoneal descending technique. In all, 87 patients underwent the operation; in two, the procedure was converted to open surgery. Peri-operative parameters which showed improvement included: operating time, blood loss, resort to blood transfusions, and the complication rate. There was no operation-related mortality. In organ-confined disease, a clear surgical margin was achieved in 93% of the patients, but in those whose disease was not organ-confined, the positive margin rate was 87%. Among patients with organ-confined disease, 13% had evidence of biochemical recurrence. Hormonal therapy was started in five patients, none of whom died during the follow-up period (mean, 24 months). Continence recovered in 69% of the patients by 6 months and in 92% by 12 months post-surgery. Assessment of erectile function before and after the surgery was problematic and estimated to be 20% among patients having the nerve-sparing procedure performed. CONCLUSION: Although Hong Kong has a relatively low incidence for prostate cancer, it was possible to develop laparoscopic radical prostatectomy with acceptable early results. Further follow-up is warranted before formulating definitive conclusions about this procedure.


Assuntos
Laparoscopia , Prostatectomia/métodos , Neoplasias da Próstata/cirurgia , Idoso , Hong Kong/epidemiologia , Humanos , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Complicações Pós-Operatórias , Neoplasias da Próstata/epidemiologia , Neoplasias da Próstata/patologia , Estudos Retrospectivos , Resultado do Tratamento
6.
Hong Kong Med J ; 13(4): 311-3, 2007 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-17592176

RESUMO

Ten young ketamine abusers presented with lower urinary tract symptoms to two regional hospitals in Hong Kong. Investigations demonstrated contracted bladders and other urinary tract abnormalities. These types of findings have never been reported before in ketamine abusers. The possible aetiology is also discussed.


Assuntos
Ketamina/intoxicação , Transtornos Relacionados ao Uso de Substâncias/complicações , Bexiga Urinaria Neurogênica/induzido quimicamente , Adulto , Feminino , Humanos , Masculino , Bexiga Urinaria Neurogênica/diagnóstico
7.
Biomaterials ; 24(17): 2821-9, 2003 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-12742720

RESUMO

Amorphous carbon films have attracted much attention recently due to their good biocompatibility. Diamond-like carbon (DLC), one form of amorphous carbon that is widely used in many kinds of industries, has been proposed for use in blood contacting medical devices. However, the blood coagulation mechanism on DLC in a biological environment is not well understood. Platelet adhesion and activation are crucial events in the interactions between blood and the materials as they influence the subsequent formation of thrombus. In this work, the behavior of platelets adhered onto hydrogenated amorphous carbon films (a-C:H) is investigated. Hydrogenated amorphous carbon films with different hydrogen contents, structures, and chemical bonds were fabricated at room temperature using plasma immersion ion implantation-deposition (PIII-D). The wettability of the films was investigated by contact angle measurements using several common liquids. Platelet adhesion experiments were conducted to examine the interaction of blood with the films in vitro and the activation of adherent platelets. The results show that the behavior of the platelets adhered on the a-C:H films is influenced by their structure and chemical bond, and it appears that protein interaction plays a key role in the activation of the adherent platelets.


Assuntos
Plaquetas/fisiologia , Plaquetas/ultraestrutura , Carbono/química , Materiais Revestidos Biocompatíveis/química , Teste de Materiais , Ativação Plaquetária/fisiologia , Contagem de Células , Células Cultivadas , Materiais Revestidos Biocompatíveis/síntese química , Diamante/química , Hidrogenação , Adesividade Plaquetária/fisiologia , Propriedades de Superfície , Tensão Superficial , Molhabilidade
8.
DNA Seq ; 12(1): 85-9, 2001 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-11697148

RESUMO

A cDNA, designated LNX1, has been identified by subtractive hybridization on the basis that it is expressed in normal human prostate but not in LNCaP cells. Sequence analysis revealed that it contained two Alu repetitive sequences but no long open reading frame. Hence, it belongs to a class of untranslated Alu-containing RNAs. LNX1 is expressed in most tissues. It is encoded by a single copy gene, which is localized on human chromosome 14.


Assuntos
Elementos Alu , Regiões não Traduzidas/genética , Sequência de Bases , Cromossomos Humanos Par 14/genética , Clonagem Molecular , DNA Complementar/genética , Feminino , Expressão Gênica , Humanos , Masculino , Dados de Sequência Molecular , Gravidez , Distribuição Tecidual
9.
DNA Cell Biol ; 20(6): 349-57, 2001 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-11445006

RESUMO

A cDNA, designated PTX1, has been isolated by subtractive hybridization on the basis that it is expressed in normal prostate but not in prostate carcinoma. The full-length cDNA was subsequently established by 5' and 3' RACE. Nucleotide sequence analysis of the 5'- and 3'-RACE clones yielded a composite cDNA of 1327 bp, which predicted a protein of 377 amino acid residues with a putative nuclear import signal (RRLNRKK) at its N terminus. The PTX1 gene was localized to human chromosome 12 and was found to be ubiquitously expressed. A segment of the cDNA was expressed in E. coli to produce a fragment of the PTX1 protein for the generation of specific antibodies. The resulting antibodies detected a 73-kDa protein in both nuclear and cytoplasmic extracts of prostate, although the level in the cytoplasmic extract was much lower. Using immunohistochemical analysis, the PTX1 protein was localized mainly in the nuclei of glandular epithelia of normal prostate. The nuclear staining was greatly reduced in prostate carcinoma. The gene organization of PTX1 was established by comparing the cDNA sequence with the published human genomic sequence.


Assuntos
Regulação Neoplásica da Expressão Gênica , Proteínas de Neoplasias/genética , Proteínas Nucleares/genética , Neoplasias da Próstata/genética , Sequência de Aminoácidos , Sequência de Bases , Western Blotting , Cromossomos Humanos Par 12 , Clonagem Molecular , DNA Complementar , DNA de Neoplasias , Escherichia coli , Humanos , Imuno-Histoquímica , Masculino , Dados de Sequência Molecular , Proteínas de Neoplasias/biossíntese , Proteínas Nucleares/biossíntese , Próstata/metabolismo , Neoplasias da Próstata/metabolismo , Alinhamento de Sequência , Proteínas de Transporte Vesicular
10.
Mol Reprod Dev ; 57(4): 361-5, 2000 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-11066065

RESUMO

Tissue distribution and cellular localization of PC1/3 mRNA in porcine tissues were examined by ribonuclease protection assay and in situ hybridization. PC1/3 mRNA was detected mainly in the corpus luteum of pregnant sow and brain. Within the ovary, PC1/3 and relaxin transcripts colocalized within large luteal cells. Levels of PC1/3 transcripts in corpora lutea increased as gestation advanced, parallel with an observed increase in relaxin transcripts. A role for PC1/3 in proprotein processing in the ovary is discussed.


Assuntos
Ácido Aspártico Endopeptidases/genética , Ovário/metabolismo , Prenhez , Animais , Ácido Aspártico Endopeptidases/metabolismo , Feminino , Expressão Gênica , Gravidez , Pró-Proteína Convertases , Suínos , Distribuição Tecidual
11.
Biopolymers ; 47(1): 101-23, 1998.
Artigo em Inglês | MEDLINE | ID: mdl-9692331

RESUMO

The solution to the protein folding problem lies in defining the relative energetic contributions of short-range and long-range interactions. In other words, the tendency of a stretch of amino acids to adopt a final secondary structural fold is context dependent. Our approach to this problem is to address whether an amino acid sequence, a "cassette," with a defined secondary structure in the three-dimensional structure of a native protein, can adopt a different conformation when placed into a different protein environment. Thus, we designed de novo a disulfide-bridged two-stranded alpha-helical parallel coiled coil, where each polypeptide chain consisted of 39 residues, as a "cassette holder." The 11-residue cassette would be inserted into the center of each polypeptide chain between the two nucleating alpha-helices to replace the control sequence. This Structural Cassette Mutagenesis model permits the analysis of short-range interactions within the inserted cassette as well as long-range interactions between the nucleating helices and the cassette region. The cassette holder, with a control sequence as the cassette, had a GdnHCl transition midpoint during denaturation of 5.6M. To demonstrate the feasibility of our model, an 11-residue beta-strand cassette from an immunoglobulin fold was inserted. The cassette was fully induced into the alpha-helical conformation with a [GdnHCl]1/2 value of 3.2M. To demonstrate the importance of short-range interactions (beta-sheet/alpha-helical propensities of amino acid side chains) in modulating structure and stability, a series of 1-5 threonine residues (highest beta-sheet propensity) were substituted into the solvent-exposed portions of the cassette in the alpha-helical conformation. Each successive substitution systematically decreased the stability of the coiled coil with peptide T4b (4 Thr residues) having a [GdnHCl]1/2 value of 2.2M. The single substitution of Ile in the hydrophobic core of the cassette with Ala or Thr had the most dramatic effect on protein stability (peptide 120T, [GdnHCl]1/2 value of 1.4M). Though these substitutions were able to modulate stability, they were not able to disrupt the alpha-helical conformation of the cassette, showing the importance of the nucleating alpha-helices on either side of the cassette in controlling conformation of the cassette. We have demonstrated the feasibility of our model protein to accept a beta-strand cassette. The effect of cassettes containing other beta-strands, beta-turns, loops, regions of undefined structure, and helical segments on conformation and stability of our model protein will also be determined.


Assuntos
Mutagênese Insercional , Engenharia de Proteínas , Sequência de Aminoácidos , Biopolímeros/química , Desenho de Fármacos , Estabilidade de Medicamentos , Humanos , Cadeias lambda de Imunoglobulina/química , Modelos Moleculares , Dados de Sequência Molecular , Peptídeos/síntese química , Peptídeos/química , Dobramento de Proteína , Estrutura Secundária de Proteína , Termodinâmica
12.
Biol Reprod ; 59(1): 202-7, 1998 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-9675013

RESUMO

The porcine seminal plasma protein (PSP) accounts for much more than 50% of the total proteins in seminal plasma. PSP has been previously purified and its biochemical properties characterized. However, the biological functions of PSP remain to be elucidated. We hypothesize that PSP is involved in the regulation of uterine immune activity. In the current study, effects of PSP on in vitro lymphocyte activities and the presence of PSP binding sites on lymphocytes were examined. In mitogen-induced proliferation assay, lymphocytes from peripheral blood of gilts were cultured with pokeweed mitogen (PWM), phytohemagglutinin (PHA), or concanavalin A (Con A) in the presence or absence of PSP. PSP at 50, 125, and 250 ng/well augmented PWM-induced [3H]thymidine uptake in a dose-responsive manner by 152.8 +/- 8.1%, 225.9 +/- 35.2%, and 274.8 +/- 53.6%, respectively, compared with that of control. PSP did not alter lymphocyte proliferation in the absence of PWM. Similarly, PSP had little or no effect on PHA- or Con A-induced lymphocyte proliferation. In one-way mixed lymphocyte reactions, PSP at 50, 125, and 250 ng/well enhanced [3H]thymidine uptake in a dose-responsive manner by 181.5 +/- 16.5%, 339.9 +/- 48.2%, and 600.1 +/- 84.8% of control, respectively. Using biotinylated PSP-I, PSP binding sites were localized on approximately 3-5% of the lymphocyte population. In summary, we have demonstrated that PSP itself is not a mitogen/antigen to porcine lymphocytes but that it has a stimulatory effect on lymphocyte activities initiated by PWM or surface antigens of lymphocytes. PSP may exert its functions by interacting with PSP binding sites on a subpopulation of porcine lymphocytes. The high potency of PSP on lymphocyte activities and the abundance of PSP in seminal plasma have suggested that PSP may play an important role in regulating immune responses in the porcine uterine environment.


Assuntos
Linfócitos/efeitos dos fármacos , Linfócitos/imunologia , Proteínas Secretadas pela Próstata , Proteínas/farmacologia , Animais , Sítios de Ligação , Biotinilação , Células Cultivadas , Concanavalina A/farmacologia , DNA/biossíntese , Feminino , Ativação Linfocitária/efeitos dos fármacos , Masculino , Mitose , Fito-Hemaglutininas/farmacologia , Mitógenos de Phytolacca americana/farmacologia , Proteínas/isolamento & purificação , Proteínas de Plasma Seminal , Suínos
13.
J Reprod Fertil ; 114(1): 77-84, 1998 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-9875158

RESUMO

Pig seminal proteins PSP-I and PSP-II are major protein components of boars' ejaculate and are present as heterodimers (PSP-dimer) in seminal plasma. These proteins were examined for their ability to modulate pig lymphocyte activity in vitro in mitogen-induced lymphocyte proliferation assays and in one-way mixed lymphocyte reactions. Pig lymphocytes were cultured with phytohaemagglutinin, concanavalin A, or pokeweed mitogen (PWM) in the presence or absence of pig seminal proteins and the amount of cellular [3H]thymidine was used as an indication of proliferation. In the absence of mitogens, none of the three pig seminal proteins affected lymphocyte proliferation suggesting that these proteins are not antigenic or mitogenic. PSP-dimer enhanced lymphocyte proliferation induced by PWM (156-227%, P < 0.05) in a concentration-dependent manner, but had no effect on phytohaemagglutinin- or concanavalin A-induced proliferation. PSP-I enhanced (127-185%, P < 0.05) phytohaemagglutinin-induced proliferation. PSP-II augmented (130-240%, P < 0.05) lymphocyte proliferation induced by concanavalin A and PWM. Lymphocytes from gilts were significantly more responsive to concanavalin A- and PWM-induced lymphocyte proliferation in the presence of PSP-I compared with boars (concanavalin A: gilts 131%, boars 91%; PWM: gilts 188%, boars 134%; P < 0.05). In the mixed lymphocyte reaction, pretreating stimulating cells with increasing concentrations of PSP-I or PSP-II elicited a 400% concentration-dependent increase (P < 0.01) in lymphocyte proliferation. The abundance of pig seminal proteins in boar seminal plasma, their ability to enhance lymphocyte proliferation, and their previously reported ability to bind to lymphocytes suggest that these proteins are immunostimulatory and supports the hypothesis that they modulate uterine immune activity to ensure reproductive success.


Assuntos
Glicoproteínas/farmacologia , Linfócitos/efeitos dos fármacos , Proteínas Secretadas pela Vesícula Seminal , Suínos/imunologia , Animais , Divisão Celular/efeitos dos fármacos , Células Cultivadas , Concanavalina A/farmacologia , Relação Dose-Resposta a Droga , Feminino , Masculino , Fito-Hemaglutininas/farmacologia , Mitógenos de Phytolacca americana/farmacologia
14.
Endocrinology ; 137(11): 5009-19, 1996 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-8895375

RESUMO

The rat placental PRL family consists of proteins structurally related to pituitary PRL. As a consequence of attempting to characterize the gene for one of the members of the family, PRL-like protein-C (PLP-C), we identified a related gene that we have termed PLP-C variant (PLP-Cv). In this study, we present information on the PLP-Cv gene and its pattern of expression. Screening of a rat genomic library with a PLP-C cDNA resulted in the isolation of four phage clones. Nucleotide sequence analysis of the clones revealed a gene, PLP-Cv, closely related but distinct from PLP-C. The PLP-Cv gene possessed a six exon/five intron organization, unique among members of the PRL family, and was localized to chromosome 17 of the rat genome, similar to other PRL family members. A PCR strategy involving primers based on the PLP-Cv gene was used to isolate a placental PLP-Cv cDNA. PLP-Cv showed 90 and 78% sequence identity with PLP-C at nucleotide and amino acid levels, respectively. Expression of PLP-Cv was restricted to the trophoblast lineage and was coordinately activated with PLP-C beginning at day 11 of gestation and continuing until term. Primer extension analysis revealed multiple putative transcription start sites. A 2.1-kilobase pair PLP-Cv promoter-luciferase reporter construct was specifically activated in differentiating rat trophoblast cells but not in other cell types. In conclusion, we have identified a new member of the PRL family possessing considerable homology to PLP-C, a unique gene structure, and displaying a trophoblast-specific pattern of transcriptional activation.


Assuntos
Mapeamento Cromossômico , Decídua/metabolismo , Proteínas da Gravidez/biossíntese , Proteínas da Gravidez/genética , Trofoblastos/metabolismo , Sequência de Aminoácidos , Animais , Sequência de Bases , Linhagem Celular , Primers do DNA , DNA Complementar , Éxons , Feminino , Expressão Gênica , Biblioteca Genômica , Células Híbridas , Camundongos , Dados de Sequência Molecular , Reação em Cadeia da Polimerase/métodos , Gravidez , Proteínas da Gravidez/química , Regiões Promotoras Genéticas , Ratos , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/química , Reprodutibilidade dos Testes , Mapeamento por Restrição , Homologia de Sequência de Aminoácidos , Transcrição Gênica
15.
Endocrinology ; 137(11): 5020-7, 1996 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-8895376

RESUMO

In this report, we have investigated placental lactogens (placental lactogen-I, PL-I; PL-I variant, PL-Iv; PL-II) expressed by differentiated Rcho-1 trophoblast cells. A complementary DNA (cDNA) library to differentiated Rcho-1 trophoblast cells was constructed and screened with probes to detect PL-I and PL-II. Sequence analysis of three independent Rcho-1 PL-I cDNAs indicated that they significantly differed from the previously reported PL-I sequence but more closely resembled a related cDNA referred to as PL-I mosaic (PL-Im). Upon further analysis, Rcho-1 PL-I/PL-Im transcripts could be detected in Rcho-1 trophoblast cells and normal developing placental tissue; however, the previously reported PL-I transcript could not be identified from the same sources. Given these results, we examined the original PL-I cDNA by PCR and nucleotide sequence analyses. The sequence differed from the original report and was found to be identical to the Rcho-1 PL-I and PL-Im cDNA clones. Thus, PL-I, Rcho-1 PL-I, and PL-Im are equivalent and should be referred to as PL-I. The PL-I gene was localized to chromosome 17 of the rat genome, similar to other PRL family members. Rcho-1 PL-II cDNAs were identical to the published PL-II sequence. PL-Iv cDNAs were isolated from differentiated Rcho-1 cells via an RT-PCR strategy and found to be identical to previously isolated PL-Iv cDNAs. Rcho-1 PL-I and PL-II cDNAs were subcloned into the pcDNA3 expression vector and recombinant protein produced in HRP-1 cells. Both recombinant Rcho-1 PL-I and PL-II proteins significantly stimulated the proliferation of lactogen-dependent rat Nb2 lymphoma cells and mouse mammary epithelial cells. In summary, we show that the Rcho-1 PL-I corresponds to PL-Im and Rcho-1 PL-Iv and PL-II are identical to their previously described placental counterparts. Additionally, both recombinant Rcho-1 PL-I and PL-II proteins are biologically active.


Assuntos
Mapeamento Cromossômico , Placenta/metabolismo , Lactogênio Placentário/biossíntese , Trofoblastos/metabolismo , Animais , Sequência de Bases , Linhagem Celular , Primers do DNA , DNA Complementar , Feminino , Expressão Gênica , Biblioteca Gênica , Variação Genética , Células Híbridas , Camundongos , Camundongos Endogâmicos BALB C , Mosaicismo , Lactogênio Placentário/genética , Reação em Cadeia da Polimerase , Gravidez , Ratos
16.
Mol Cell Endocrinol ; 116(1): 49-58, 1996 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-8822264

RESUMO

Placenta lactogen-I variant (PL-Iv) is a member of a family of proteins expressed by the rat placenta with characteristics similar to prolactin (PRL). In this report, we present the molecular cloning, chromosomal localization, and heterologous expression of PL-Iv. Nucleotide sequence analysis of the PL-Iv cDNA clone predicted a precursor protein of 223 amino acids, including a 28-amino acid signal sequence. The PL-Iv gene was localized to chromosome 17 of the rat genome, which also carries other members of the PRL gene family. PL-Iv heterologously expressed in Chinese Hamster ovary (CHO) cells exhibited similar immunoreactive and electrophoretic characteristics with PL-Iv produced by the rat placenta. N-terminal sequencing verified the identity and purity of the recombinant PL-Iv species and the site of cleavage of the signal peptide from the mature secreted PL-Iv species. Recombinant PL-Iv was shown to bind to ovarian and liver PRL receptors, stimulate the proliferation of Nb2 lymphoma cells, and activate Jak2. Each of these actions is consistent with PL-Iv utilizing the PRL receptor signal transduction pathway.


Assuntos
Lactogênio Placentário/genética , Lactogênio Placentário/metabolismo , Receptores da Prolactina/metabolismo , Animais , Mapeamento Cromossômico , Clonagem Molecular , Cricetinae , DNA Complementar/genética , Feminino , Expressão Gênica , Variação Genética , Gravidez , Sinais Direcionadores de Proteínas/genética , Sinais Direcionadores de Proteínas/metabolismo , Ratos , Transdução de Sinais
17.
Biochim Biophys Acta ; 1264(1): 1-6, 1995 Oct 17.
Artigo em Inglês | MEDLINE | ID: mdl-7578241

RESUMO

Overlapping cDNAs encoding porcine prohormone convertase, PC1/3, have been isolated from a pregnant sow ovary cDNA library using a mouse PC1/3 cDNA as a probe. Nucleotide sequence analysis of these cDNAs predicts a PC1/3 precursor protein of 753 amino acid residues, which shares an overall sequence homology of 96, 92, and 92% with the human, rat, and mouse counterparts, respectively. Furthermore, five different polyadenylation sites have been observed. The utilization of these polyadenylation sites results in a length difference of 40-440 bp in the 3' untranslated regions of the transcripts.


Assuntos
Ácido Aspártico Endopeptidases/genética , Ovário/enzimologia , Pró-Proteína Convertase 1 , Sequência de Aminoácidos , Animais , Sequência de Bases , Sítios de Ligação , Sondas de DNA , DNA Complementar/química , DNA Complementar/isolamento & purificação , Feminino , Dados de Sequência Molecular , Gravidez , Pró-Proteína Convertases , Homologia de Sequência de Aminoácidos , Suínos
18.
DNA Cell Biol ; 13(4): 389-94, 1994 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-8011165

RESUMO

A full-length cDNA encoding the porcine acrosin inhibitor has been isolated from a boar seminal vesicle cDNA library. Nucleotide sequence analysis of the 667-bp cDNA predicts a precursor protein of 97 amino acid residues, which includes a 26-residue signal peptide and a 71-residue secreted protein. The predicted amino acid sequence of the mature protein agrees completely with that of the sperm-associated acrosin inhibitor determined by conventional amino acid sequence analysis. However, the asparagine/aspartic acid and glutamine/glutamic acid substitutions, as reported in the seminal plasma counterpart, have not been observed. Southern blot analysis shows only a single hybridizing band with three different restriction endonucleases, suggesting the presence of a single copy of the acrosin inhibitor gene in the porcine genome.


Assuntos
Acrosina/antagonistas & inibidores , Proteínas Secretadas pela Vesícula Seminal , Inibidor da Tripsina Pancreática de Kazal/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Northern Blotting , Southern Blotting , Clonagem Molecular , DNA Complementar , Humanos , Masculino , Dados de Sequência Molecular , Precursores de Proteínas/genética , Homologia de Sequência de Aminoácidos , Suínos , Distribuição Tecidual , Inibidor da Tripsina Pancreática de Kazal/metabolismo
19.
J Biol Chem ; 269(9): 6517-23, 1994 Mar 04.
Artigo em Inglês | MEDLINE | ID: mdl-8120003

RESUMO

The purpose of this study was to investigate P450scc expression during trophoblast differentiation. Biochemical characteristics of P450scc protein and mRNA identified in rat trophoblast tissues were similar to those identified in the rat adrenal gland. Furthermore, P450scc was localized to trophoblast giant cells. This observation prompted an examination of progesterone biosynthesis and P450scc expression in Rcho-1 cells. Rcho-1 cells were derived from a transplantable rat choriocarcinoma, their differentiation can be regulated, and they have the capacity to express the trophoblast giant cell phenotype. Progesterone was produced by Rcho-1 cells and increased approximately 100-fold as the cells progressed from proliferation to differentiation. P450scc protein and mRNA accumulation also increased during trophoblast differentiation. P450scc expression within the Rcho-1 cell line was restricted to trophoblast giant cells. To further investigate the regulation of P450scc expression during trophoblast differentiation, we examined a plasmid construct, containing 894 base pairs of DNA 5' upstream from the P450scc transcriptional start site linked to a human growth hormone reporter gene, following stable transfection into Rcho-1 cells. The transfected P450scc regulatory DNA permitted the expression of human growth hormone which paralleled expression of the endogenous P450scc gene. In conclusion, transcriptional activation of the P450scc gene accompanies trophoblast giant cell differentiation.


Assuntos
Enzima de Clivagem da Cadeia Lateral do Colesterol/biossíntese , Regulação Enzimológica da Expressão Gênica , Trofoblastos/citologia , Trofoblastos/enzimologia , Alantoide/enzimologia , Animais , Diferenciação Celular , Linhagem Celular , Células Cultivadas , Enzima de Clivagem da Cadeia Lateral do Colesterol/análise , Enzima de Clivagem da Cadeia Lateral do Colesterol/genética , Córion/enzimologia , Feminino , Microscopia Imunoeletrônica , Placenta/enzimologia , Gravidez , Progesterona/biossíntese , RNA Mensageiro/biossíntese , RNA Mensageiro/metabolismo , Ratos , Fatores de Tempo , Transcrição Gênica , Transfecção , Trofoblastos/ultraestrutura
20.
DNA Cell Biol ; 12(7): 605-10, 1993 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-8397818

RESUMO

Two full-length cDNAs encoding porcine seminal proteins, PSP-I and PSP-II, have been isolated from a porcine seminal vesicle cDNA library. Nucleotide sequence analysis of the 706-bp PSP-I cDNA predicts a precursor protein of 133 amino acid residues, which includes a 21-residue signal peptide and a 112-residue secreted protein. On the other hand, the complete sequence of the 686-bp PSP-II cDNA reveals a coding sequence for a 21-residue signal peptide and a 116-residue secreted protein. The predicted amino acid sequences agree very well with those determined by conventional amino acid sequence analysis. Alignment of the two cDNA sequences shows an overall 66% sequence homology throughout their entire length. However, the sequence homology is much higher in the 3' untranslated region (72%) than in the coding region (61%). This suggests that these two genes evolved by duplication and divergence from a common ancestral gene. They share about 50% amino acid sequence homology and a similar overall structure with three members of the spermadhesin family.


Assuntos
Glicoproteínas/genética , Proteínas Secretadas pela Vesícula Seminal , Sequência de Aminoácidos , Animais , Sequência de Bases , Clonagem Molecular , Dados de Sequência Molecular , Precursores de Proteínas/genética , Homologia de Sequência de Aminoácidos , Homologia de Sequência do Ácido Nucleico , Suínos
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