Antinutritional factors and hypocholesterolemic effect of wild apricot kernel (Prunus armeniaca L.) as affected by detoxification.

The present investigation was aimed to study the effect of detoxification on the nutrients and antinutrients of wild apricot kernel followed by its hypocholesterolemic effect in male Wistar albino rats. The results revealed a non-significant (p > 0.05) effect of detoxification on the proximate composition except total carbohydrates and protein content. However, detoxification led to a significant (p < 0.05) decrease in l-ascorbic acid (76.82%), ß-carotene (25.90%), dietary fiber constituents (10.51-28.92%), minerals (4.76-31.08%) and antinutritional factors (23.92-77.05%) (phenolics, tannins, trypsin inhibitor activity, saponins, phytic acid, alkaloids, flavonoids, oxalates) along with the complete removal (100%) of bitter and potentially toxic hydrocyanic acid (HCN). The quality parameters of kernel oil indicated no adverse effects of detoxification on free fatty acids, lipase activity, acid value and peroxide value, which remained well below the maximum permissible limit. Blood lipid profile demonstrated that the detoxified apricot kernel group exhibited significantly (p < 0.05) increased levels of HDL-cholesterol (48.79%) and triglycerides (15.09%), and decreased levels of total blood cholesterol (6.99%), LDL-C (22.95%) and VLDL-C (7.90%) compared to that of the raw (untreated) kernel group. Overall, it can be concluded that wild apricot kernel flour could be detoxified efficiently by employing a simple, safe, domestic and cost-effective method, which further has the potential for formulating protein supplements and value-added food products.

The effects of osterix on the proliferation and odontoblastic differentiation of human dental papilla cells.

INTRODUCTION: Dental papilla cells (DPCs) are precursors of odontoblasts and have the potential to differentiate into odontoblasts. Osteoblasts and odontoblasts have many common characteristics. Osterix (Osx) is essential for osteoblast differentiation. However, no information is available for the effects of Osx on the odontoblastic differentiation of DPCs. The purpose of this study was to investigate the effects of Osx on the proliferation and odontoblastic differentiation of DPCs. METHODS: An immortalized human dental papilla cell (hDPC) line was used. Osx was stably overexpressed or knocked down in hDPCs with infection of lentiviral particles to determine its biological effects on hDPCs. The proliferation of cells was measured by the 5-ethynyl-2'-deoxyuridine incorporation assay and direct cell counting. Expressions of dentin sialophosphoprotein, nestin, dentin matrix protein 1, and alkaline phosphatase were detected by real-time polymerase chain reaction to determine the odontoblastic differentiation of cells. The mineralization ability of cells was evaluated by von Kossa staining and alkaline phosphatase activity assay. RESULTS: Overexpression of Osx retarded the proliferation of hDPCs, whereas knockdown of Osx increased the cell proliferation. Overexpression of Osx promoted the odontoblastic differentiation of hDPCs by up-regulating odontoblastic differentiation genes and increased the mineralization ability of hDPCs. Knockdown of Osx down-regulated odontoblastic differentiation genes and decreased the mineralization ability of hDPCs. CONCLUSIONS: Osx might function as a potential regulator for the proliferation and odontoblastic differentiation of hDPCs.

Variability in tannin content, chemistry and activity in a diverse group of tannin containing sorghum cultivars.

BACKGROUND: Tannins are large polyphenolic polymers and are known to bind proteins, limiting their digestibility, but are also excellent antioxidants. Numerous studies investigating the functional properties of sorghum tannin have been conducted by comparing grain samples from different sorghum lines without considering the other intrinsic characteristics of the grain. The purpose of this study was to remove the confounding intrinsic factors present in the endosperm so the effect of the tannins could be evaluated utilizing a unique decortication/reconstitution procedure. RESULTS: The tannin content of the 14 cultivars tested ranged from 2.3 to 67.2 catechin equivalents. The bran fractions were studied for their impact on protein binding and antioxidant capacity. Protein digestibility by pepsin ranged from 8% to 58% at the highest tannin level addition. Protein binding ranged from 3.11 to 16.33 g blue bovine serum albumin kg⁻¹ bran. Antioxidant capacity ranged from 81.33 to 1122.54 µmol Trolox equivalents g⁻¹ bran. High-performance size-exclusion chromatography detailed molecular size distributions of the tannin polymers and relationship to tannin functionality. CONCLUSION: The tannin content and composition play a significant role in determining tannin functionality. These differences will allow for selections of high-tannin sorghums with consideration of the biological activities of the tannins.

Optimization of process for reduction of antinutritional factors in edible cereal brans.

Reduction of various antinutritional factors in cereal brans by different treatments (microwave heating, dry heating and wet heating) were studied. There was significant difference (p ≤ 0.05) in reduction of antinutritional factors of treated cereal brans except for dry heating at low temperature. Microwave heating at 2450 MHz for 2.5 min resulted in 53.85%, 57.21%, 65.00% and 100% reduction in phytic acid, polyphenols, oxalates and saponins, respectively. Wet heating resulted in maximum reduction in trypsin inhibitor activity (83.07%) at 110 °C for 25 min. Processing treatment resulted in increase in bulk density and slight darkening of the brans. The most effective method of detoxifying most of the toxicants was microwave heating for 2.5 min, and therefore it could be exploited for making treated brans an ideal source for potential food application.

Determination of propylthiouracil in pharmaceutical formulation by high-performance liquid-chromatography with a post-column iodine-azide reaction as a detection system.

A high-performance liquid chromatographic method with a post-column iodine-azide reaction has been chosen and tested for validity in quantitative determination of propylthiouracil in tablets. A mobile phase with a flow rate of 1.4 ml/min was conducted in the form of isocratic chromatography on a C18 column with acetonitrile-water-sodium azide solution (2.5%; pH 5.5) 24:26:50 (v/v/v). Unreacted iodine from post-column iodine-azide induced by reaction was monitored with visible detection at lambda=350 nm. The method proved both its linearity within the range of 8-100 nM (r2>0.9988) and satisfactory results of inter-day precision (RSD<4.2%) and accuracy (recovery>91%). The limits of detection (DDL) and quantification (DQL) reached the levels of 5 and 8 nM, respectively. The validation of the method comprised also its specificity. The results obtained proved the suitability and appropriateness of the suggested method for intended use.

Ambient air pollution and preterm birth in the environment and pregnancy outcomes study at the University of California, Los Angeles.

The authors conducted a case-control survey nested within a birth cohort and collected detailed risk factor information to assess the extent to which residual confounding and exposure misclassification may impact air pollution effect estimates. Using a survey of 2,543 of 6,374 women sampled from a cohort of 58,316 eligible births in 2003 in Los Angeles County, California, the authors estimated with logistic regression and two-phase models the effects of pregnancy period-specific air pollution exposure on the odds of preterm birth. For the first trimester, the odds of preterm birth consistently increased with increasing carbon monoxide exposures and also at high levels of exposure to particulate matter less than or equal to 2.5 microm in diameter (>21.4 microg/m(3)), regardless of type of data (cohort/sample) or covariate adjustment (carbon monoxide exposures of >1.25 ppm increased the odds by 21-25%). Women exposed to carbon monoxide above 0.91 ppm during the last 6 weeks of pregnancy experienced increased odds of preterm birth. Crude and birth certificate covariate-adjusted results for carbon monoxide differed from each other. However, further adjustment for risk factors assessed in the survey did not change effect estimates for short-term pollutant averages appreciably, except for time-activity patterns, which strengthened the observed associations. These results confirm the importance of reducing exposure misclassification when evaluating the effect of traffic-related pollutants that vary spatially.

Ex vivo application of carbon monoxide in University of Wisconsin solution to prevent intestinal cold ischemia/reperfusion injury.

Carbon monoxide (CO), a byproduct of heme catalysis, was shown to have potent cytoprotective and anti-inflammatory effects. In vivo recipient CO inhalation at low concentrations prevented ischemia/reperfusion (I/R) injury associated with small intestinal transplantation (SITx). This study examined whether ex vivo delivery of CO in University of Wisconsin (UW) solution could ameliorate intestinal I/R injury. Orthotopic syngenic SITx was performed in Lewis rats after 6 h cold preservation in control UW or UW that was bubbled with CO gas (0.1-5%) (CO-UW). Recipient survival with intestinal grafts preserved in 5%, but not 0.1%, CO-UW improved to 86.7% (13/15) from 53% (9/17) with control UW. At 3 h after SITx, grafts stored in 5% CO-UW showed improved intestinal barrier function, less mucosal denudation and reduced inflammatory mediator upregulation compared to those in control UW. Preservation in CO-UW associated with reduced vascular resistance (end preservation), increased graft cyclic guanosine monophosphate levels (1 h), and improved graft blood flow (1 h). Protective effects of CO-UW were reversed by ODQ, an inhibitor of soluble guanylyl cyclase. In vitro culture experiment also showed better preservation of vascular endothelial cells with CO-UW. The study suggests that ex vivo CO delivery into UW solution would be a simple and innovative therapeutic strategy to prevent transplant-induced I/R injury.

Trypan blue identifies antimetabolite treatment area in trabeculectomy.

AIM: Colourless solutions of mitomycin C (MMC) and 5-fluorouracil (5-FU) are widely used during trabeculectomy to inhibit postoperative scarring. The poor visibility of these agents on the eye has several drawbacks including the inability to accurately assess the area of treatment. This study examined the utility of using trypan blue dye to colour antimetabolites used during trabeculectomy and the effect of trypan blue on antimetabolite cytotoxicity in vitro. METHODS: For in vitro experiments, MMC (0.4 mg/ml) and 5-FU (25 mg/ml) were reconstituted with or without trypan blue. A lactate dehydrogenase release assay was used to measure drug induced cell death and viable cell number 7 days after treatment. For clinical assessment, trypan blue 0.1% was added to MMC and 5-FU to final concentrations of between 0.01% and 0.05%. The mixture was applied to Tenon's capsule and sclera via pre-wet or into dry 5x8 mm sponges (MMC and 5-FU) for 3 minutes or by direct subconjunctival injection after completion of surgery (5-FU). Twenty two consecutive patients undergoing trabeculectomy either with or without trypan blue were followed for 2 years postoperatively. RESULTS: The addition of 0.05% trypan blue to MMC or 5-FU did not alter MMC induced cell death or the number of viable fibroblast in vitro. In vivo, trypan blue clearly delineated the antimetabolite treatment area and facilitated control of excess antimetabolite at the wound margins as well as sponge removal. With direct subconjunctival injection, total staining area varied for a given volume with location of the needle tip. Any leakage from the injection site could be easily seen. No adverse effects attributable to trypan blue were found in 2 years of follow up. CONCLUSIONS: Trypan blue permits delineation of antimetabolite/tissue interactions without affecting cytoxicity for the assays investigated. Trypan blue can be used to visualise antimetabolite soaked sponges, estimate treatment area, and show areas of unintended tissue contact during trabeculectomy. The addition of trypan blue to antimetabolites has potential benefits in clinical, research, and teaching aspects of ocular surgery and therapy.

Hydrophilic interaction liquid chromatography-APCI-mass spectrometry determination of 5-fluorouracil in plasma and tissues.

A simple and fast analytical method using hydrophilic interaction liquid chromatography (HILIC) coupled with mass spectrometry was developed to analyse 5-fluorouracil (5-FU) in plasma and tissues. The HILIC system overcomes problems reported in obtaining satisfactory retention of 5-FU with other types of HPLC systems. After addition of internal standard (IS) (5-Chlorouracil (5-CU)), plasma proteins were precipitated with acetonitrile, and tissue samples homogenised with a micro-dismembrator. The analysis was performed using a polymer-based column (Ashaipak NH2) and the compounds were eluted under gradient conditions at 1 ml/min using a mobile phase containing a mixture of ammonium formate and acetonitrile. MS detection used a API 4000 mass spectrometry with heated nebulizer source and multiple reaction monitoring operated in the negative ion mode. The mass transitions of 5-FU and its internal standard were 129 m/z-->42m/z and 145 m/z-->42 m/z, respectively. The lower limits of quantitation in plasma and tissues were about 5 ng/ml and 10 ng/g, respectively, using 25 microl of plasma and 50mg of tissue. Good linearity, accuracy and precision were obtained in all matrices tested. The suitability and robustness of the method for in vivo samples were confirmed by analysis of mouse plasma, muscle and tumour from animals dosed with 5-FU.

EPR studies of 5-bromouracil crystal after irradiation with X rays in the bromine K-edge region.

Radicals induced in a single crystal of 5-bromouracil (BrUra) by synchrotron soft X rays in the bromine K-edge region (13.461-13.482 keV) were investigated using the X-band EPR method. The crystal was irradiated at three peak energies of the absorption spectrum at room temperature or at 80 K. A hydrogen abstraction radical derived from N1 of the pyrimidine ring was commonly observed for all of the energies used, though with some variation in quantity. Similar characteristics were also observed in the EPR signal for the off-K-edge low-energy (13.42 keV) and (60)Co gamma rays used for comparison. When irradiated at 80 K, a much larger exposure (roughly 10 times) of soft X rays was needed to obtain the same signal intensity as that observed at room temperature. EPR signals were not detectable with gamma irradiation at liquid nitrogen temperature.

Time of origin of the rat pineal gland cells. A bromodeoxyuridine immunohistochemical study.

The immunohistochemical detection of bromodeoxyuridine (BrdU) was used to study the time of origin of the cells in the pineal gland of the rat. A study was made involving 17 groups of 4 rats each, administered with a single dose of bromodeoxyuridine (BrdU, 25 mg/kg) in 7 phases of the embryonic period (E15 to E21) and in 10 postnatal phases (between P0 and P30), followed by determination in each rat of the number of visible immune-labeled cells in the pineal gland 60 days after birth. The results show that approximately 60% of the pineal cells underwent the last division(s) prior to differentiation in the prenatal period between E18 and E21. The rest of the pineal cells originated after birth, particularly in the first 5 postnatal days.

Hypothermia inhibits cell proliferation and nitric oxide synthase expression in rats.

In the present study, the effects of cold-water immersion on cell proliferation and nitric oxide synthase expression in the dentate gyrus of rats were investigated. Sprague-Dawley rats were divided into four groups: the control-rest group; the control-heat group; the cold-rest group; and the cold-heat group. Cold-water immersion for 5 min at 4 degrees C suppressed the numbers of 5-bromo-2'-deoxyuridine-positive and nicotinamide adenine dinucleotide phosphate-diaphorase-positive cells in the dentate gyrus, and these numbers were increased by warming for 30 min at 30 degrees C. In the present study, it was demonstrated that warming protects against cold stress-induced suppression of new cell formation, and results suggest that nitric oxide, the synthesis of which is affected adversely by cold-water immersion, may play an important role in the regulation of cell proliferation.

Electrospray ionization mass spectrometry and tandem mass spectrometry of clodronate and related bisphosphonate and phosphonate compounds.

The bisphosphonate family with a P-C-P structure is a broad class of drugs, widely investigated as potential inhibitors in bone diseases and calcium metabolic disorders. In this study, the mass spectrometric (MS) behavior and fragmentation of clodronate and related bisphosphonate and phosphonate compounds was studied by using negative ion electrospray ionization (ESI) with triple quadrupole and ion trap instruments. The effect of pH on the degree of deprotonation of the polyprotic bisphosphonic and phosphonic acids in negative ion ESI-MS was investigated, and the degree of deprotonation in the ESI mass spectra and the dissociation in the liquid phase were compared. The results provide evidence that the measured ESI mass spectra do not correlate with the chemistry in the liquid phase owing to the decrease in the pH of the solvent droplets during the ion evaporation process and the charge state neutralization in the gas phase. Ion trap MS(n) provided useful information on the fragmentation study of clodronate and related bisphosphonate and phosphonate compounds, in which interesting fragmentation pathways including the direct elimination of carbon monoxide from deprotonated bisphosphonates and formation of a P-P bond were observed. Reactions between the product ions with a -PO(2) group and residual water in the ion trap or in the high-pressure region of the triple quadrupole instrument formed other unexpected fragmentation paths for all the bisphosphonates studied.

Suppression of proliferative cholangitis by E2F decoy oligodeoxynucleotide.

BACKGROUND: Proliferative cholangitis (PC) associated with hepatolithiasis results in stricture of the main bile ducts and is a major cause of residual and/or recurrent stones after repeated treatment for hepatolithiasis. The transcription factor E2F controls the expression of several genes involved in cell proliferation. The aim of this study was to inhibit PC using cytostatic gene therapy by transferring fusigenic anionic liposome-hemagglutinating virus of Japan (HVJ-anionic liposome) complexes containing a synthetic double-stranded oligodeoxynucleotide with high affinity for E2F (E2F decoy). MATERIALS AND METHODS: PC was induced by introducing a fine nylon thread into the bile duct in a rat model. HVJ-anionic liposomes containing the E2F decoy were administered directly into the biliary tract. HVJ-anionic liposomes containing a missense oligodeoxynucleotide (scramble decoy) were also given as a control. The count of peribiliary glands in the bile duct, 5'-bromodeoxyuridine (BrdU) labeling index, and immunohistochemical staining for proliferating cell nuclear antigen (PCNA) in the bile duct were compared among untransfected, scramble decoy-transfected, and E2F decoy-transfected rats. RESULTS: E2F decoy-transfected bile ducts showed inhibition of the papillary proliferation of the biliary epithelium and peribiliary gland hyperplasia. BrdU incorporation and PCNA expression in the bile ducts were inhibited in E2F decoy-transfected rats. CONCLUSION: Our cytostatic gene therapy approach using direct E2F decoy transfer into the biliary tract suppressed PC in a rat model and may offer an effective therapeutic option for reducing recurrence following treatment for hepatolithiasis.

Schwann cell-enriched cultures from adult human peripheral nerve: a technique combining short enzymatic dissociation and treatment with cytosine arabinoside (Ara-C).

Attempts to design the nerve cellular prostheses have focused on the production of autologous Schwann cells expanded in vitro as the essential component in the regeneration process of injured peripheral nerves. To obtain human Schwann cells of high quality we tested a short enzymatic dissociation protocol that optimized cellular viability levels. We also assessed patterns of bromodeoxyuridine (BrdU) incorporation in both Schwann cells and fibroblasts in the presence or absence of the antimitotic Ara-C, an enrichment option for adult human Schwann cell cultures. The Ara-C treated cultures showed a significantly higher Schwann cell percentage (95%), compared with that obtained in the absence of Ara-C (70%), indicating that this antimitotic acts to eliminate fibroblasts in each one of the applied pulses (four pulses). However, we have observed that the use of this antimitotic during prolonged periods of time produced a cumulative effect causing Schwann cell cytotoxicity. Therefore, we consider that our enzymatic dissociation technique and the application of only two pulses of Ara-C to the cultures are enough to achieve enrichment of adult human Schwann cells in culture.

Sexually dimorphic neurogenesis is topographically matched with the anterior accessory olfactory bulb of the adult rat.

The accessory olfactory bulb (AOB) is a sexually dimorphic structure of the vomeronasal system, which plays a role in the control of sexual behaviors. In adult rats, we have demonstrated previously that the migrating neuroblasts of the subependymal layer (SEL) directed to the main olfactory bulb (MOB) also reach the AOB. To tackle the relation between sexual dimorphism and targeted cell migration, we quantified the neo-neurogenesis in the AOB of adult rats of both sexes. Our results confirm a morphological sexual dimorphism in the AOB granular layer volumes. We showed that the number of newly generated cells reaching the AOB in both sexes was considerable, even if lower than those directed to the MOB. Moreover, we demonstrated that the rate of neurogenesis in the anterior AOB of the two sexes was significantly different.

A detailed analysis of cyclin A accumulation at the G(1)/S border in normal and transformed cells.

The temporal relationship between cyclin A accumulation and the onset of DNA replication was analyzed in detail. Five untransformed and nine transformed asynchronously growing cell cultures were investigated using a triple immunofluorescence staining protocol combined with computerized evaluation of staining intensities in individual cells. The simultaneous staining of BrdU, cyclin A, and cyclin E made it possible to determine the cell cycle position of each cell investigated. Cells at the G(1)/S border were identified on the basis of cyclin E content and were further analyzed with respect to cyclin A and BrdU content. A method was developed to calculate objective thresholds defining the highest staining intensity found in the negative cells in the population. Using the thresholds we could distinguish cells with minute amounts of cyclin A and BrdU from truly negative cells. We show that the onset of cyclin A accumulation and the start of DNA replication occurs at the same time, or deviating by a few minutes at the most. We also show that cyclin A accumulates continuously during S. This study clearly demonstrates that nuclear cyclin A can be used as a reliable marker for the S and G(2) phases in both normal and transformed interphase cells.

Cell damage and neurogenesis in the dentate granule cell layer of adult rats after pilocarpine- or kainate-induced status epilepticus.

Dentate granule cells are generally considered to be relatively resistant to excitotoxicity and have been associated with robust synaptogenesis after neuronal damage. Synaptic reorganization of dentate granule cell axons, the mossy fibers, has been suggested to be relevant for hyperexcitability in human temporal lobe epilepsy and animal models. A recent hypothesis suggested that mossy-fiber sprouting is dependent on newly formed dentate granule cells. However, we recently demonstrated that cycloheximide (CHX) can block the mossy-fiber sprouting that would otherwise be induced by different epileptogenic agents and does not interfere with epileptogenesis in those models. Here, we investigated cell damage and neurogenesis in the dentate gyrus of pilocarpine- or kainate-treated animals with or without coadministration of CHX. Dentate granule cells were highly vulnerable to pilocarpine induced-status epilepticus (SE), but were hardly damaged by kainate-induced SE. CHX pretreatment markedly reduced the number of injured neurons after pilocarpine-induced SE. Induction of SE dramatically increased the mitotic rate of KA- and KA + CHX-treated animals. Induction of SE in animals injected with pilocarpine alone led to 2-7-fold increases in the mitotic rate of dentate granule cells as compared to 5- and 30-fold increases for pilocarpine + CHX animals. We suggest that such increased mitotic rates might be associated with a protection of a vulnerable precursor cell population that would otherwise degenerate after pilocarpine-induced SE. We further suggest that mossy-fiber sprouting and neurogenesis of granule cells are not necessarily linked to one another.

Olfactory epithelium consisting of supporting cells and horizontal basal cells in the posterior nasal cavity of mice.

The olfactory epithelium of mice generally consists of olfactory cells, progenitors of olfactory cells (globose basal cells), supporting cells, and horizontal basal cells. However, in the dorsal fossa (the roof) of the posterior nasal cavity of mice, we found seven epithelial patches consisting of only non-neuronal cell types, i.e., supporting cells and horizontal basal cells, among the normal olfactory epithelium. The supporting cells occupied three or four layers in the apical to middle regions; in the basal region, horizontal basal cells were localized in a single row adjacent to the basement membrane. Bowman's gland ducts were also present in the epithelium. Neuronal cells (olfactory cells and globose basal cells) were totally absent. The ultrastructure of the supporting cells, horizontal basal cells, and Bowman's glands was essentially similar to that in the normal olfactory epithelium. In the early postnatal period (P1-P7), cell types in the epithelium were the same as those in the normal olfactory epithelium. From P10 to P21, olfactory cells and globose basal cells had disappeared from the olfactory epithelium. At this period, the number of TUNEL-positive cells was significantly higher than that in the surrounding olfactory epithelium; ultrastructurally, many apoptotic figures were observed. This suggests that the epithelium consisting of supporting cells and horizontal basal cells is generated by the apoptotic death of olfactory cells and globose basal cells during postnatal development.

Protein phosphatase inhibitors arrest cell cycle and reduce branching morphogenesis in fetal rat lung cultures.

Protein phosphatase 2A (PP2A) is a key signal transduction intermediate in the regulation of cellular proliferation and differentiation in vitro. However, the role of PP2A in the context of a developing organ is unknown. To explore the role of PP2A in the regulation of lung development, we studied the effect of PP2A inhibition on new airway branching, induction of apoptosis, DNA synthesis, and expression of epithelial marker genes in whole organ explant cultures of embryonic (E14) rat lung. Microdissected lung primordia were cultured in medium containing one of either two PP2A inhibitors, okadaic acid (OA, 0-9 nM) or cantharidin (Can, 0-3,600 nM), or with the PP2B inhibitor deltamethrin (Del, 0-10 microM) as a control for a PP2A-specific effect for 48 h. PP2A inhibition with OA and Can significantly inhibited airway branching and overall lung growth. PP2B inhibition with Del did not affect lung growth or new airway development. Histologically, both PP2A- and PP2B-inhibited explants were similar to controls. Increased apoptosis was not the mechanism of decreased lung growth and new airway branching inasmuch as OA-treated explant sections subjected to the terminal deoxynucleotidyltransferase dUTP nick end labeling reaction demonstrated a decrease in apoptosis. However, PP2A inhibition with OA increased DNA content and 5-bromo-2'-deoxyuridine uptake that correlated with a G(2)/M cell cycle arrest. PP2A inhibition also resulted in altered differentiation of the respiratory epithelium as evidenced by decreased mRNA levels of the early epithelial marker surfactant protein C. These findings suggest that inhibition of protein phosphatases with OA and Can halted mesenchymal cell cycle progression and reduced branching morphogenesis in fetal rat lung explant culture.