Correlation between amniotic fluid lamellar body concentrations at different gestational ages and respiratory distress syndrome

Background: Respiratory distress syndrome (RDS) also known as hyaline membrane disease is the most common cause of respiratory failure in neonates. The risk of RDS rises in prematurity due to decreased production of surfactant. Lamellar bodies (LB) are storage form of surfactant and are actively secreted into the alveolar space and hence into the amniotic fluid. The objective of the present study was to find out correlation between amniotic fluid lamellar body concentrations (LBC) at different gestational ages and respiratory distress syndromeMethods: The study was a prospective observational study conducted over a period of 1 year at department of OBG, SDM college of medical sciences, Dharwad. All women undergoing caesarean section were included in the study. 5 ml of amniotic fluid was collected at the time of Caesarean section and sent to laboratory, Lamellar body concentrations (LBC) was counted in auto analyzer by platelet impedance counting. Incidence of RDS at different gestational age with an LBC cut off 40,500 was calculated.Results: Among 300 patients studied, Respiratory distress was seen in 116 (41.3%) of patients. LBC cut off of 40,500 was chosen. Out of 140 preterm babies, 121 (86.8%) had lamellar body count less than 40,500 and 19 (13.5%) had more than 40,500. There is significant correlation between decreasing lamellar body count in preterms and increasing incidence of RDS.Conclusions: LBC count increases with increasing gestational ages and is inversely proportional to the incidence of RDS in preterm newborns.

Correlation between ABCA3 and Pediatric Pulmonary Disease / 临床儿科杂志

Pulmonary surfactant is a lipid-protein complex that lines and stabilizes the respiratory interface in the alveoli, allowing for gas exchange during the breathing cycle. ATP-binding Cassette transporters A3, is a lipid transporter in the limiting membrane of lamellar bodies in alveolar type Ⅱ cells that plays a critical role in the regulation of pulmonary surfactant homeostasis. Mutations in the ABCA3 gene cause respiratory distress syndrome in new-born and childhood interstitial lung disease. An updated view on expression of ABCA3 gene and ABCA3 mutation associated with neonatal respiratory distress syndrome,or childhood interstitial lung disease was reviewed.

Amniotic Fluid Lamellar Body Count as a Predictor of Fetal Lung Maturity.

Assessment of fetal lung maturity can be done with various biochemical tests on amniotic fluid. In the present study, lamellar body count (LBC) was compared with the traditional shake bubble test (SBT), as LBC is a simple, reliable and cost-effective method to predict fetal lung maturity. This was a prospective study conducted on 50 patients with known last menstrual period (LMP), gestational age >28 weeks presenting in active phase of labor with intact membranes. Amniotic fluid was collected either through vaginal route or through amniocentesis and SBT and LBC were conducted on all samples. Newborns were watched for signs of respiratory distress syndrome (RDS). Fourteen babies developed RDS. i.e., one out of 34 babies with positive SBT, seven out of 10 with intermediate SBT, all six with negative SBT developed RDS. The LBC varied between 3,000-28,000/ml among the 14 babies with RDS. i.e., all cases with RDS had LBC <30,000/ml. This study showed that LBC with cut-off value of 30,000/ml to predict fetal lung maturity is superior to all other tests in terms of technique and cost-effectiveness.

Lamellar Body Counts in Fetal Rabbits' Experimental Diaphragmatic Hernia and Tracheal Ligation

Experimental tracheal ligation (TL) has been shown to reverse the pulmonary hypoplasia associated with congenital diaphragmatic hernia (CDH) and to normalize gas exchange. The purpose of this study was to determine whether the TL would correct the surfactant deficiency present in the fetal rabbit model of CDH by using lamellar body count. Lamellar bodies are synthesized and secreted by the type II pneumocytes of fetal lung. The phospholipids present in these bodies constitute the major component of pulmonary surfactant. Twenty-one pregnant New Zealand rabbits underwent hysterotomy and fetal surgery on gestational day 24. Two fetuses of each pregnant rabbit were operated. In the fetus of one end of bicornuate uterus, left DH was created by excision of fetal diaphragm through open thoracotomy (DH Group). In the fetus of the other end of bicornuate uterus, left DH and TL were created (TL Group). The fetuses were delivered by Cesarean section on gestational day 31. Fourteen in control group, 12 in the DH group and 13 in TL group were born alive. En bloc excision of lungs, bronchi and trachea was done in all newborn rabbits. A five Fr catheter was inserted through trachea and repeated irrigations with 10 cc normal saline were done. The irrigated fluid was centrifuged at 280 xg for 5 minutes and the lamellar bodies were counted with the upper level fluid in platelet channel of electronic cell counter. The average lamellar body counts were 37.1 +/- 14.2 x 10(3)/microL in control group, 11.5 +/- 4.4 x 10(3)/microL in DH group, and 6.5+/- 0.9 x 10(3)/microL in TL group. Lamellar body count in DH group was lower than in control group and did not increase after TL. This study shows TL has no therapeutic effect on decreased surfactant level of CDH and the pregnant rabbit is appropriate for the animal model of CDH.

Lamellar body distribution, ceramidase expression and skin barrier function in lesions of polymorphous light eruption / 中华皮肤科杂志

Objective To investigate the correlation of epidermal distribution of lamellar bodies and expression of ceramidase with skin barrier dysfunction in polymorphous light eruption.Methods Forty-seven patients with polymorphous light eruption and 40 healthy volunteers were recruited into this study.Noninvasive instruments were used to measure skin sebum content,transepidermal water loss(TEWL)and water content in stratum corneum in all of the subjects.Then,tissue specimens were obtained from the lesions at sunexposed sites in the patients and normal skin of the healthy volunteers.The ultrastructure and distribution of lamellar bodies were observed with transmission electron microscopy in five lesion and control specimens.Immunohistochemistry was performed to detect the expression of ceramidase in the tissue specimens.Results Compared with the normal skin from healthy volunteers,the lesions from patients showed decreased number of lamellar bodies in the granular layer and prick cell layer with a disorganized arrangement.Ceramidase was positively expressed in 20 lesion specimens and 36 normal control specimens,weakly expressed in 21 lesion specimens and 4 normal control specimens,and negative in 6 lesion specimens; there was a significant difference in the expression of ceramidase between the lesion specimens and normal control specimens(P ＜ 0.01).The lesions also showed high TEWL(34.2191 ± 12.70 vs.16.8350 ± 6.50,P ＜ 0.01),lower water content in stratum corneum(22.7319 ± 8.71 vs.29.4250 ± 5.08,P ＜ 0.01)and similar skin sebum content compared with the normal skin.Conclusions There is a disturbance in the synthesis of ceramide in patients with polymorphous light eruption,which may contribute to the impairment of skin barrier.

Change of alveolar epithelial type Ⅱ cells and pulmonary surfactant protein A in young rats with acute lung injury / 中华急诊医学杂志

Objective To study the temporal changes of alveolar epithelial type Ⅱ cells and surfactant pro-tein A in young rats with acute lung injury induced by lipopolysaecharide. Method Totally 110 SD young rats (male:53, female : 57) were randomly divided into ALI and normal control groups (six subgroups in each group).LPS(4 mg/kg) was given intraperitoneally in ALI group. The same amount of normal saline was given in the con-trol groups. Eight rats in each subgroup were sacrificed at 6, 12, 24, 36, 48 and 72 hours after the injection.Lung samples were taken for transmission electron microscope examination. RT-PCR was epmloyed for the mea-surement of SP-A mRNA. Western blot was used for the detection of SP-A in the lung tissue. ANOVA and homo-geneity of variance test were performed by SPSS 12.0. Results The microvilli disappeared at 24 hours after the injection of LPS. The number of lamellar body (LBs) was provisionality increased at 24 hours and 48 hours. The ring-like an'angement of LBs around nucleus and the giant LB with vacuole-like deformity were found as the main characteristics of AEC- Ⅱ in ALI at 48 hours. The number of LBs reduced and broken and residual LB remained at 72 hours. SP-A elevated greatly from 24 to 48 hours (P < 0.01), reached peak at 36 hours (6.94 ± 0.80, P <0.01),reached the lowest level(3.87 ±0.50, P <0.01)at 72 hours. Conclusions The pathological changes of AEC-Ⅱ and SP-A in lung tissue wiht ALI are time-dependent. The typical alterations of AEC- Ⅱ occurs at 48 hours accompanied by the compensatory increase of SP-A. AEC- Ⅱ is seriously injuried with the typical changes of LBs and the diminishing of SP-A in lung tissue.

An Update of the Defensive Barrier Function of Skin

Skin, as the outermost organ in the human body, continuously confronts the external environment and serves as a primary defense system. The protective functions of skin include UV-protection, anti-oxidant and antimicrobial functions. In addition to these protections, skin also acts as a sensory organ and the primary regulator of body temperature. Within these important functions, the epidermal permeability barrier, which controls the transcutaneous movement of water and other electrolytes, is probably the most important. This permeability barrier resides in the stratum corneum, a resilient layer composed of corneocytes and stratum corneum intercellular lipids. Since the first realization of the structural and biochemical diversities involved in the stratum corneum, a tremendous amount of work has been performed to elucidate its roles and functions in the skin, and in humans in general. The perturbation of the epidermal permeability barrier, previously speculated to be just a symptom involved in skin diseases, is currently considered to be a primary pathophysiologic factor for many skin diseases. In addition, much of the evidence provides support for the idea that various protective functions in the skin are closely related or even co-regulated. In this review, the recent achievements of skin researchers focusing on the functions of the epidermal permeability barrier and their importance in skin disease, such as atopic dermatitis and psoriasis, are introduced.

An Update of the Defensive Barrier Function of Skin

Skin, as the outermost organ in the human body, continuously confronts the external environment and serves as a primary defense system. The protective functions of skin include UV-protection, anti-oxidant and antimicrobial functions. In addition to these protections, skin also acts as a sensory organ and the primary regulator of body temperature. Within these important functions, the epidermal permeability barrier, which controls the transcutaneous movement of water and other electrolytes, is probably the most important. This permeability barrier resides in the stratum corneum, a resilient layer composed of corneocytes and stratum corneum intercellular lipids. Since the first realization of the structural and biochemical diversities involved in the stratum corneum, a tremendous amount of work has been performed to elucidate its roles and functions in the skin, and in humans in general. The perturbation of the epidermal permeability barrier, previously speculated to be just a symptom involved in skin diseases, is currently considered to be a primary pathophysiologic factor for many skin diseases. In addition, much of the evidence provides support for the idea that various protective functions in the skin are closely related or even co-regulated. In this review, the recent achievements of skin researchers focusing on the functions of the epidermal permeability barrier and their importance in skin disease, such as atopic dermatitis and psoriasis, are introduced.

Epidermal Barrier in Human Fetus and Newborn / 대한피부과학회지

BACKGROUND: The lipids of the stratum corneum, which originate from polar lipid precursors provided by the cells of the stratum granulosum via the exocytosis of lamellar bodies, with cornified cell envelope form competent epidermal barrier structurally and functionally. The ontogeny of the epidermal barrier is not clearly defined because of difficulty of sampling and methodology which defines epidermal lipids. OBJECT: From ultrastructural observation of skin samples obtained from human fetuses and newborn on serial developmental timings, we tried to clarify the sequential development of epidermal barrier. METHODS: Skin samples were obtained from 13 human fetuses from EGA(estimated gestational age) 10 to 23wks and 2 newborns. Specimens were observed by fluorescent confocal microscopy with nile red to identify the distribution of epidermal lipids, by transmission electron microscope with lanthanum to investigate the functional permeability barrier, with RuO4 to observe the intercellular lipid bilayer and morphology of lamellar bodies, with ion capture cytochemistry to investigate the formation of epidermal calcium gradient. RESULT: In nile red stain, the amount of epidermal lipid increased during fetal period. At EGA 23wks, the lipid distribution revealed linear and continuous pattern. In lanthanum tracer study, the electron dense tracer permeated all the intercellular space of the epidermis up to periderm and subepidermal space until EGA 21wks. At EGA 23wks, the tracer permeated intercellular space of epidermis weakly. It might be predicted that incomplete epidermal barrier is present at this time. In RuO4 stain, precursor of lamellar body was observed at EGA 15wks, and intercellular lipid bilayer was observed at EGA 16wks. As gestation increases, there was a steady increase in epidermal lipid bilayers. In ion capture cytochemistry, epidermal calcium gradient was first observed in follicular epidermis at EGA 20wks, and in interfollicular epidermis at EGA 23wks. From these results, it is concluded that the basic structures of epidermal barrier are formed at EGA 23wks, but it is not complete, and epidermal barrier arises first from follicular epidermis.

The Efficacy of Lamellar Body Counts for Prediction of Perinatal Outcome / 대한산부인과학회잡지

OBJECTIVE: This study is aimed to evaluate whether lamellar body count (LBC) in amniotic fluid could be used as a predictor of neonatal morbidity as well as respiratory distress syndrome (RDS) and to determine the value of lamellar body count that maximizes sensitivity and specificity. METHODS: We conducted a prospective clinical outcome study. Amniotic fluid was obtained from 39 pregnant women at various gestational ages (29 to 36 weeks) from March 1, 2002 to February 28, 2003. They delivered within 72 hours of amniocentesis, excluding 6 cases of contaminated amniotic fluid. A LBC was performed on each specimen. The frequencies of RDS, minor and major morbidity of newborn in different LBC cutoff value were analyzed. Statistic analysis was done by Chi-square test. RESULTS: LBCs increased with gestation (r=0.533, p<0.05). The LBC cutoff value that best agreed with RDS (sensitivity 36.4%, specificity 93.8%), minor morbidity (sensitivity 100.0%, specificity 50.0%) and major morbidity (sensitivity 45.5%, specificity 81.3%) was 30,000/ l. CONCLUSION: These results indicate that LBC might be useful as a predictor of neonatal morbidity as well as respiratory distress syndrome.

The clinical efficacy and cost-effectiveness of Lamellar body count in fetal lung maturation test / 대한산부인과학회잡지

OBJECTIVE: To evaluate the availability and efficacy of the Lamellar body count as a predictor of fetal lung maturity METHODS: Amniocentesis was performed for evaluation of fetal lung maturity status within 72 hours of delivery in 32 patients. A Lamellar body count in clean amniotic fluid (AF) was analyzed right after amniocentesis. In case of contaminated AF with meconium or blood, samples was centrifuged for 3 minutes and divided into three. The average of three samples was analyzed successively from Coulter counter. RESULTS: The incidence of RDS showed significant relation with gestational age, 1-minute and 5-minute Apgar Score. Using a value greater than 30,000/ l to indicate pulmonary maturity, the Lamellar body count predicted all lung-maturated cases with no false-negative results (100% negative predictive value). All 4 cases of RDS demonstrated Lamellar body count less than 30,000/microl. By using a lower cutoff of 10,000/microl to predict pulmonary immaturity. Positive predictive value was 100% in RDS patients. CONCLUSION: In high risk pregnancy, fetal lung maturity test from amniotic fluid lamellar body count is a rapid, simple and reliable method in making a decision of delivery-time. And also it has universal availability with cost-effectiveness when we consider the reality of korean medical situation

The Clinical Efficacy of Lamellar Body Count to Predict Fetal Lung Maturity / 대한산부인과학회잡지

From July 1994 through August 1995, 31 patients who delivered before 37 weeks gestation at the Ajou University Hospital were tested for lamellar body count in the amniotic fluid. The obtained results are summarized as follows; 1. Lamellar body count was associated with 1 minute Apgar score(p=0.02). 2. There was no sexual, gestational and birth weight differences in the incidence of respiratory distress syndrome. 3. Lamellar body count showed that the positive predictive value for respiratory distress syndrome was 90.0 %, the negative predictive value was 95.2 %, and the sensitivity was 90.0 %, and the specificity was 95.2 %, therefore it may be regarded as more accurate than other previously employed methods. This study suggests that the lamellar body count reflects fetal lung maturity sufficiently to assure that the neonate will not develop respiratory distress syndrome, and can be used as a quick screening test or one in series of tests for thorough assessment of fetal lung maturity.

Pulmonary Alveolar Proteinosis: A case report

A case of pulmonary alveolar proteinosis is reported. Most of the alveolar spaces were filled with amorphous deep eosinohilic material which revealed strong positive reaction to periodic acid-Schiff staining. Electron microscopic observation of this material showed numerous lamellar bodies in the alveolar spaces and cytoplasms of alveolar macrophages. A part of them were concentric multilamellated type A lamellar bodies and the other were finger printlike type B bodies. Combined type A and type B lamellar bodies were rarely present. From the above features it is suggested that both type A and B lamellar bodies could be transformed one another and those lamellar bodies may be originated from pulmonary surfactant.

Relationship between the phospholipid content of BALL and lamellar body in septic rats / 中国病理生理杂志

The phospholipid content in BALL markedly increased (P0.01) in septic rats induced by cecal ligation plus punctures as compared with non-septic control. Meanwhile, the lamellar inclusion bodies in type Ⅱ alveolar epithelial cells were empty observed in electro-microscopic examination. Authors suggest that the changes mentioned above is a compensatory response related to stress in early stage of sepsis.

A STEREOLOGIC ANALYSIS OF LAMELLAR BODY WITHIN PULMONARY ALVEOLAR TYPE Ⅱ CELLS IN DOGS WITH ACUTE HEMORRHAGIC PANCREATITIS / 第二军医大学学报

A stereologic study of lamellar bodies within pulmonary alveolar type II cells in dogs with experimental acute hemorrhagic pancreatitis(group P)and in dogs inje- cted muscularly with choloroquine before inducing pancreatitis (group PI)was carried out.Our findings showed that volumerical density (Vv), surface density (Sv)and numerical density(Nv)of lamellar bodies were decreased by 29.06%,30.28% and 40.75% in the group P as compared with those in the control group (group N), respectively, and their Sv and Nv were decreased by 23.8% and 30.26% in comparison with those in the group PI.The results suggest that the lung injury resulting from acute hemorrhagic pancreatitis was characterized by alveolar surfactant System damage.Dramatical decrease in the number of the lamellar bodies affected the surface tension of alveoli in a certain degree, leading to atelectasis.There was a relationship between lung injury and the effect of PLA2(phospholipidase A2).Choloroquine might play a role in maintaining the number of lamellar bodies.