The impact of postpartum post-traumatic stress disorder symptoms on child development: a population-based, 2-year follow-up study.

BACKGROUND: Against the background of very limited evidence, the present study aimed to prospectively examine the impact of maternal postpartum post-traumatic stress disorder (PTSD) symptoms on four important areas of child development, i.e. gross motor, fine motor, communication and social-emotional development. METHOD: This study is part of the large, population-based Akershus Birth Cohort. Data from the hospital's birth record as well as questionnaire data from 8 weeks and 2 years postpartum were used (n = 1472). The domains of child development that were significantly correlated with PTSD symptoms were entered into regression analyses. Interaction analyses were run to test whether the influence of postpartum PTSD symptoms on child development was moderated by child sex or infant temperament. RESULTS: Postpartum PTSD symptoms had a prospective relationship with poor child social-emotional development 2 years later. This relationship remained significant even when adjusting for confounders such as maternal depression and anxiety or infant temperament. Both child sex and infant temperament moderated the association between maternal PTSD symptoms and child social-emotional development, i.e. with increasing maternal PTSD symptom load, boys and children with a difficult temperament were shown to have comparatively higher levels of social-emotional problems. CONCLUSIONS: Examining four different domains of child development, we found a prospective impact of postpartum PTSD symptoms on children's social-emotional development at 2 years of age. Our findings suggest that both boys and children with an early difficult temperament may be particularly susceptible to the adverse impact of postpartum PTSD symptoms. Additional studies are needed to further investigate the mechanisms at work.

Efficacy of an emollient cream in the treatment of xerosis in diabetic foot: a double-blind, randomized, vehicle-controlled clinical trial.

BACKGROUND: Peripheral neuronal impairment compromises foot health in patients with diabetes. Clinically, xerosis is the most common mild complication, but it should not be underestimated. An effective treatment must be able to restore the cutaneous barrier and prevent water loss, to maintain adequate hydration and protection. OBJECTIVE: This study aimed to assess the efficacy of an emollient cream on foot xerosis in patients with diabetes. METHODS: This is a prospective, multicenter, randomized, double-blind contralateral vehicle-controlled study in 57 patients with diabetes. Patients were treated twice daily for 27 ± 2 days with the study emollient containing glycerol 15%, liquid and soft paraffin 10%, glycerol monostearate, stearic acid, polydimethylcyclosiloxane, silicone oil, macrogol 600, trolamine, propyl parahydroxybenzoate and purified water (Dexeryl® ; Pierre Fabre Medicament, Boulogne, France) or its vehicle (glycerol monostearate, stearic acid, polydimethylcyclosiloxane, silicone oil, macrogol 600, trolamine, propyl parahydroxybenzoate and purified water). Efficacy was assessed after a 28-day treatment period using a validated score [Xerosis Assessment Scale (XAS) score], instrumental measurements and subjective assessment. RESULTS: The XAS score decreased to 3.2 ± 2.6 points with the emollient and 4.1 ± 2.3 with the vehicle (P = 0.001). Improvement was observed from day 14 (P = 0.012). Compared with the vehicle, the emollient also significantly improved the overall skin score, hydration index, D-Squame® (CuDerm Corporation, Dallas, TX, USA) test, skin roughness and patients' opinions. CONCLUSION: Treatment with an emollient is effective for improving foot xerosis in patients with diabetes.

Peripartum changes in social support among women with and without anxiety and depressive disorders prior to pregnancy: a prospective-longitudinal study.

This study aims to prospectively examine peripartum changes in social support in women with and without anxiety and depressive disorders prior to pregnancy. Data come from the Maternal Anxiety in Relation to Infant Development (MARI) Study, a prospective-longitudinal investigation among n = 306 expectant mothers. DSM-IV anxiety and depressive disorders were assessed in early pregnancy using the Composite International Diagnostic Interview for Women (CIDI-V). Social support was assessed with the Social Support Questionnaire during pregnancy as well as 4 and 16 months postpartum. Perceived social support in the total sample declined from prepartum to postpartum. Levels of prepartum and postpartum social support were lower in women with comorbid anxiety and depressive disorders compared to those with pure depressive disorder(s), pure anxiety disorder(s), or comorbid anxiety and depressive disorders prior to pregnancy. Moreover, social support more strongly declined from prepartum to postpartum in women with comorbid anxiety and depressive disorders compared to those without anxiety and depressive disorder prior to pregnancy. Findings suggest that women with a previous history of comorbid anxiety and depressive disorders are at particular risk for deficient social support during pregnancy and after delivery and might thus profit from targeted early interventions.

Peripartum changes in partnership quality among women with and without anxiety and depressive disorders prior to pregnancy: a prospective-longitudinal study.

The purpose of this study is to prospectively examine peripartum changes in partnership characteristics among women with and without anxiety and depressive disorders prior to pregnancy. In the prospective-longitudinal Maternal Anxiety in Relation to Infant Development (MARI) study, n = 306 expectant mothers completed up to seven waves of assessment from early pregnancy until 16 months postpartum. Lifetime anxiety and depressive disorders according to the Diagnostic and Statistical Manual of Mental Disorders Fourth Edition (DSM-IV) were evaluated at baseline using the Composite International Diagnostic Interview for Women (CIDI-V, Martini et al. 2009). Partnership characteristics were assessed during pregnancy as well as 4 and 16 months postpartum using the Partnership Questionnaire (Hahlweg 1996). Linear regressions were applied to test associations between diagnostic status prior to pregnancy and peripartum partnership characteristics. Compared to women without anxiety and depressive disorders prior to pregnancy, women with comorbid anxiety and depressive disorders reported less tenderness during pregnancy, less postpartum tenderness, satisfaction, and overall partnership quality as well as a lower decrease in communication from pre- to postpartum. Women with pure depressive disorders and comorbid anxiety and depressive disorders prior to pregnancy indicated a higher increase in quarreling from pre- to postpartum. Findings suggest that women with depressive (and comorbid anxiety) disorders prior to pregnancy are at elevated risk for an unfavorable peripartum partnership development and might thus profit from targeted family interventions during this period.

Maternal anxiety versus depressive disorders: specific relations to infants' crying, feeding and sleeping problems.

BACKGROUND: Maternal depression has been associated with excessive infant crying, feeding and sleeping problems, but the specificity of maternal depression, as compared with maternal anxiety remains unclear and manifest disorders prior to pregnancy have been widely neglected. In this prospective longitudinal study, the specific associations of maternal anxiety and depressive disorders prior to, during and after pregnancy and infants' crying, feeding and sleeping problems were investigated in the context of maternal parity. METHODS: In the Maternal Anxiety in Relation to Infant Development (MARI) Study, n = 306 primiparous and multiparous women were repeatedly interviewed from early pregnancy until 16 months post partum with the Composite International Diagnostic Interview for Women (CIDI-V) to assess DSM-IV anxiety and depressive disorders. Information on excessive infant crying, feeding and sleeping problems was obtained from n = 286 mothers during postpartum period via questionnaire and interview (Baby-DIPS). RESULTS: Findings from this study revealed syndrome-specific risk constellations for maternal anxiety and depressive disorders as early as prior to pregnancy: Excessive infant crying (10.1%) was specifically associated with maternal anxiety disorders, especially in infants of younger and lower educated first-time mothers. Feeding problems (36.4%) were predicted by maternal anxiety (and comorbid depressive) disorders in primiparous mothers and infants with lower birth weight. Infant sleeping problems (12.2%) were related to maternal depressive (and comorbid anxiety) disorders irrespective of maternal parity. CONCLUSIONS: Primiparous mothers with anxiety disorders may be more prone to anxious misinterpretations of crying and feeding situations leading to an escalation of mother-infant interactions. The relation between maternal depressive and infant sleeping problems may be better explained by a transmission of unsettled maternal sleep to the fetus during pregnancy or a lack of daily structure and bedtime routine with the infant. Maternal disorders prior to pregnancy require more attention in research and clinical practice.

[Mental disorders in women: Natural course during premenstrual phases, peripartum period and perimenopause]. / Psychische Störungen in den reproduktiven Phasen der Frau: Häufigkeiten, Verlauf und Besonderheiten.

Epidemiological studies indicate sex-specific differences in prevalence rates and the natural course of mental disorders. Affective, anxiety, somatoform and eating disorders are more prevalent in women than men, whereas substance use disorders occur more commonly in men, and some disorders are equally distributed in both sexes (e. g. psychotic disorders). The aim of this review is to depict the natural course of mental disorders during the reproductive stages (premenstrual phases, peripartum period, perimenopause) in women, including also neuroendocrine features associated with the menstrual cycle, pregnancy, puerperium and perimenopause. Recommendations for sex-specific diagnostic and therapeutic procedures are provided.

[Fear of progression in parents of children with cancer: adaptation of the Fear of Progression Questionnaire and correlates]. / Progredienzangst bei Eltern krebskranker Kinder: Adaptation eines Fragebogens und Korrelate.

BACKGROUND: Fear of Progression (FoP), the fear of further disease progression, is one of the most common psychological strains of chronically ill patients and can also be found in healthy partners of cancer patients. Parents of children with cancer are also at risk of developing distinct fears that may persist after medical treatment. This study aimed to assess FoP in parents of children with cancer and to investigate relationships between FoP in parents of children with cancer and disease- and treatment-related issues, the child's current medical condition and parents' quality of life. PATIENTS: In this study 76 parents (51 mothers, 25 fathers) whose children were in inpatient treatment or follow-up care were surveyed. METHOD: The short form of the FoP Questionnaire was adapted by rephrasing the items for the parental perspective (FoP-Q-SF/PR). RESULTS: The FoP-Q-SF/PR is a short questionnaire with adequate psychometric properties (e. g. Cronbach's α=0.90) and satisfying results in terms of construct validity. Significant correlations with FoP are found for the child's current medical condition (r=0.35), time since diagnosis (r=- 0.30), parents' capacity to cope with disease-related fears (r=- 0.45) and parents' quality of life (r=- 0.55). A cut-off value of 46 points is recommended. CONCLUSION: The FoP-Q-SF/PR offers a feasible and sensitive battery to assess disease-related fears. For clinicians, evaluation of individual results can provide insight into specific problem areas for parents of children with cancer. The questionnaire is thus well suited for use in psychosocial care of families within the field of paediatric oncology.

Effect of ß-lactoglobulin A and B whey protein variants on cheese yield potential of a model milk system.

Cheese yield mainly depends on the amount and proportion of milk constituents; however, genetic variants of the proteins present in milk may also have an important effect. The objective of this research was to study the effect of the variants A and B of ß-lactoglobulin (LG) on cheese yield using a model system consisting of skim milk powder fortified with different levels of a mixture containing α-lactalbumin and ß-LG genetic variants (A, B, or A-B) in a 1:2 ratio. Fortified milk samples were subjected to pasteurization at 65 °C for 30 min. Miniature cheeses were made by acidifying (pH=5.9) fortified milk and incubating with rennet for 1h at 32 °C. The clot formed was cut, centrifuged at 2,600 × g for 30 min at 20 °C and drained for determining cheese yield. Cheese-yielding capacity was expressed as actual yield (grams of cheese curd per 100g of milk) and dry weight yield (grams of dried cheese curd per 100g of milk). Free-zone capillary electrophoresis was used for determining ß-LG A or B recovery in the curd during rennet-induced coagulation. The presence of ß-LG variant B resulted in a significantly higher actual and dried weight cheese yield than when A or A-B were present at levels ≤ 0.675% of whey protein (WP) addition. Results of free-zone capillary electrophoresis allowed us to infer that ß-LG B associates with the casein micelles during renneting, as shown by an increase in the recovery of this variant in the curd when ß-LG B was added up to a maximum at 0.45% (equivalent to 0.675% WP). In general, actual or dried weight cheese yield increased as WP addition was increased from 0.225 to 0.675%. However, when WP addition ranged from 0.675 to 0.90%, a drastic drop in cheese yield was observed. This behavior may be because an increase in the aggregation of casein micelles with a concomitant inclusion of whey protein in the gel occurs at low levels of WP addition, whereas once the association of WP with the casein micelles reach a saturation point at addition levels higher than 0.675%, rearrangements of the gel network result in larger whey expulsion and syneresis. This knowledge is expected to be useful to maximize cheese yield and optimize processing conditions during cheese and cheese analogs manufacturing.

Management of diabetic foot ulcers with a TLC-NOSF wound dressing.

OBJECTIVE: To evaluate the efficacy, tolerance and acceptability of UrgoStart Contact (Laboratoires Urgo), a new wound dressing impregnated with NOSF, as an MMP regulator in the management of neuropathic diabetic foot ulcers. METHOD: A multicentre, pilot, prospective, non-controlled open-label clinical trial. Adult patients with type 1 or 2 diabetes mellitus, who had a grade 1A (Texas classification), uninfected, neuropathic foot ulcer, 1-15cm2 in size and of 1-20 months' duration (mean 6.7 ± 5.2 months) were included in the study. The primary endpoint was the relative reduction of the wound surface area (%) at the end of the study. Secondary endpoints included rate of complete healing, and tolerability and acceptability of the dressing. The wound dressing was changed regularly at the investigator's discretion, in accordance with the wound status and exudate level. Patients were followed up every 2 weeks for a 12-week period. At each visit, patients underwent clinical assessments, and ulcer surface area was measured by planimetry and photographs. RESULTS: Thirty-four diabetic patients with a neuropathic foot ulcer were included but only 33 cases were analysed, as data were completely lost for one patient. At baseline, mean surface area was 2.7±2.4cm2. At the 12-week follow-up, the median surface area reduction was 82.7% (mean reduction 62.7 ± 49.9%) and in 10 of the 33 analysed patients (30%) the wound was healed. Only two of the seven documented local adverse events were deemed to be dressing related. According to the nursing staff, acceptability was considered very satisfactory, particularly in term of conformability and ease of use. CONCLUSION: This pilot study indicates that use of the new UrgoStart Contact dressing, combined with offloading and debridement,may help promote the healing process of the neuropathic diabetic foot ulcers, and was well tolerated and accepted.

Annual prevalence estimation of lymphatic malformation with a cutaneous component: observational study of a national representative sample of physicians.

BACKGROUND: Lymphatic malformations (LMs) represent a potentially life-threatening, rare disease of the lymphatic system characterized by development of abnormal vessels, outpouchings, or cysts filled with lymphatic fluid. There are three morphologic types of LMs based on the size of the individual cysts: macrocystic (typically > 2 cm), microcystic (generally < 2 cm), and mixed (includes aspects of both). Macrocystic LMs typically exist beneath the skin and often can involve vascular components and/or organs. Microcystic LMs often have a cutaneous component and clinically present with lymphorrhea, bleeding, pain, itching, malodor, and functional deficits. There are no treatments approved by the US Food and Drug Administration (FDA) for either macrocystic or microcystic lymphatic malformations. The totality of the epidemiologic literature for LM is limited to the incidence of the disease among various birth cohorts. This is the first nationally representative study to estimate the national managed prevalence for patients with microcystic LM or combined LM with a cutaneous component annually across physician specialties likely to manage this condition. We conducted a retrospective observational survey of a nationally representative sample of patient-care physicians in the United States most likely to manage lymphatic malformations with a cutaneous component (LMC). Once recruited, target physicians participated via an electronic questionnaire. We weighted study physician self-estimates of the number of LMC patients treated in the past 12 months to reflect the specialists' corresponding proportion in the national universe. All patient information was anonymous; no personally identifiable information was collected. RESULTS: Of the 420 physicians who visited the study website, 316 agreed to be screened and to participate (75.2% participation rate). Our survey results indicated the estimated number of unique annually managed LMC patients by target specialists is 79,920 (CI 66,600-93,250). This number corresponds to managed prevalence of 24.1 LMC patients per 100,000 population (CI 19.6/100,000-28.4/100,000). CONCLUSIONS: The study indicates that while rare, LMC affects a substantial number of people in the US (79,920) who are being managed by one or more specialists. By better understanding the prevalence of people living with LMC who require treatment, efforts to both increase disease awareness and to identify underserved populations in need of potential new treatments can be better focused.

Changes in psychopathological symptoms during pregnancy and after delivery: A prospective-longitudinal study in women with and without anxiety and depressive disorders prior to pregnancy.

BACKGROUND: The role of anxiety and depressive disorders prior to pregnancy for changes in peripartum psychopathological symptoms has not been resolved yet. METHODS: A regional-epidemiological sample of 306 women was prospectively followed in seven waves from early pregnancy until 16 months postpartum. Lifetime DSM-IV anxiety and depressive disorders were assessed at baseline with the CIDI-V. Psychopathological symptoms (somatization, obsession-compulsion, interpersonal sensitivity, depression, anxiety, hostility, phobic anxiety, paranoid ideation, and psychoticism) were measured with the BSI three times during pregnancy and three times after delivery. RESULTS: Multilevel analyses revealed that women with versus without lifetime anxiety (ß=0.22 to ß=0.32) and depressive (ß=0.24 to ß=0.34) disorders prior to pregnancy experienced higher peripartum psychopathological symptoms. All symptoms linearly decreased during pregnancy (ß=-0.02 to ß=-0.07 per month). Somatization (ß=-0.46) was lower, whereas paranoid ideation (ß=0.26) and obsession-compulsion (ß=0.21) were higher after delivery than during pregnancy. Though, obsession-compulsion linearly decreased after delivery (ß=-0.02). Lifetime anxiety disorders prior to pregnancy interacted with linear changes in anxiety (ß=-0.04) and phobic anxiety (ß=-0.05) during pregnancy. That is, only women with, but not without anxiety disorders prior to pregnancy experienced a linear decline in anxiety and phobic anxiety during pregnancy. LIMITATIONS: Lifetime anxiety and depressive disorders were assessed in early pregnancy and might be biased. CONCLUSIONS: Peripartum psychopathological symptoms are higher in women with versus without lifetime anxiety and depressive disorders prior to pregnancy, but symptom changes only slightly vary by lifetime diagnostic status.

Approximate Genome-Based Kernel Models for Large Data Sets Including Main Effects and Interactions.

The rapid development of molecular markers and sequencing technologies has made it possible to use genomic prediction (GP) and selection (GS) in animal and plant breeding. However, when the number of observations (n) is large (thousands or millions), computational difficulties when handling these large genomic kernel relationship matrices (inverting and decomposing) increase exponentially. This problem increases when genomic × environment interaction and multi-trait kernels are included in the model. In this research we propose selecting a small number of lines m(m < n) for constructing an approximate kernel of lower rank than the original and thus exponentially decreasing the required computing time. First, we describe the full genomic method for single environment (FGSE) with a covariance matrix (kernel) including all n lines. Second, we select m lines and approximate the original kernel for the single environment model (APSE). Similarly, but including main effects and G × E, we explain a full genomic method with genotype × environment model (FGGE), and including m lines, we approximated the kernel method with G × E (APGE). We applied the proposed method to two different wheat data sets of different sizes (n) using the standard linear kernel Genomic Best Linear Unbiased Predictor (GBLUP) and also using eigen value decomposition. In both data sets, we compared the prediction performance and computing time for FGSE versus APSE; we also compared FGGE versus APGE. Results showed a competitive prediction performance of the approximated methods with a significant reduction in computing time. Genomic prediction accuracy depends on the decay of the eigenvalues (amount of variance information loss) of the original kernel as well as on the size of the selected lines m.

Partial purification of neurofilament subunits from bovine brains and studies on neurofilament assembly.

The 200,000-dalton neurofilament subunit (P200) and the 160,000-dalton (P160) and 78,000-dalton (P78) neurofilament subunits were partially purified from bovine brain. Intact neurofilaments were prepared by high-speed and sucrose-zone centrifugation. The crude neurofilament was solubilized in 8 M urea solution containing pyridine, formic acid, and 2-mercaptoethanol. The solubilized neurofilament was purified by carboxymethyl (CM) cellulose column and hydroxylapatite column chromatography. The P200 was purified as separate from P160 and P78, but the P160 and P78 subunits were copurified on CM cellulose, hydroxylapatite, Bio-Gel A150m, and Sephadex G-150 column chromatography. Electron microscopy of these purified neurofilament subunits revealed the P200 subunit as a globular structure, and the P160 and P78 subunits as a rod-shaped structure extending up to 120 nm with a 8- to 12-nm width. In the presence of 200 mM KCl, 15 mM MgCl2, and 1 mM ATP, the purified subunits assembled into long filaments. Under the assembly condition, P160 and P78 subunits elongated up to 500 nm, but the longer filament formation required the presence of P200 subunits. The filaments formed in vitro were of two types: long straight filaments and intertwined knobby-type filaments. From these results, we have suggested that P160 and P78 form the neurofilament backbone structure and P200 facilitates the assembly of the backbone units into longer filaments.

[Diabetic foot: detection and prevention]. / Le pied diabétique : dépistage et prévention.

Foot care prevention programs can reduce the occurrence of foot ulcerations and amputations. The most important factor related to the development of foot ulcer is peripheral neuropathy, associated with loss of pain. Neuropathy can be associated with peripheral vascular disease and foot deformities. Five cornerstones for prevention should be respected, including regular examination of the feet and footwear, identification of high-risk patients, education of the patient and family, appropriate footwear, and treatment of nonulcerative pathology. After examination of the foot using the Semmes-Weinstein monofilament test, each patient can be assigned to one of the four risk categories to guide management. Foot care programs should be provided to high-risk categories of patients. Education plays an important role in prevention. The aim is to increase motivation and skills and enhance compliance with footwear advice. Items may include daily feet inspection and regular washing. Appropriate foot wear should protect against trauma. High-risk patients should participate in a foot care program set up by a multidisciplinary foot care team.

Daclatasvir and Sofosbuvir With or Without Ribavirin in Liver Transplant Recipients: A Single-Center Real-World Study.

BACKGROUND: Treatment with direct-acting antiviral drugs in interferon-free regimens is currently recommended for viral hepatitis C recurrence after liver transplantation. There are limited data regarding its results in this population, and no optimal treatment scheme has yet been singled out. METHODS: We report our real-world results in liver transplant (LT) recipients. All patients were hepatitis C virus (HCV) monoinfected and completed a 12-week treatment course, followed 12 weeks later by HCV polymerase chain reaction testing with 12 IU/mL sensibility. Liver fibrosis was graded with the use of biopsies taken <12 months before treatment and stratified as early (0-1) or moderate to advanced (2-4) according to the Metavir score. RESULTS: Median postoperative time was 5.2 years. Genotype 3 was found in 66.7% of the sample. The following regimens were prescribed: daclatasvir-sofosbuvir with (n = 11) or without (n = 28) ribavirin. Genotypes 1 and 3 were evenly distributed between the regimens. Sustained virologic response (SVR) was obtained in 24 out of 28 patients (85.7%) who received daclatasvir-sofosbuvir and in all patients (100%) who received daclatasvir-sofosbuvir-ribavirin (global SVR 89.7%). All patients that failed treatment had genotype 3 HCV. Fibrosis was evaluated in 79.5% of the sample: 48.4% had early and 51.6% had moderate to advanced fibrosis, for which ribavirin was more commonly prescribed (P = .001). CONCLUSIONS: The SVR rate in our LT recipients was similar to that previously reported in the literature. The addition of ribavirin to DAA treatment appears to be justified in this population.

Collaborating to Compete: Blood Profiling Atlas in Cancer (BloodPAC) Consortium.

The cancer community understands the value of blood profiling measurements in assessing and monitoring cancer. We describe an effort among academic, government, biotechnology, diagnostic, and pharmaceutical companies called the Blood Profiling Atlas in Cancer (BloodPAC) Project. BloodPAC will aggregate, make freely available, and harmonize for further analyses, raw datasets, relevant associated clinical data (e.g., clinical diagnosis, treatment history, and outcomes), and sample preparation and handling protocols to accelerate the development of blood profiling assays.

Beneficial effects due to increasing blood and plasma viscosity.

Increased plasma and blood viscosity are usually associated with pathological conditions; however there are several situations in which the elevation of both parameters results in increased perfusion and the lowering of peripheral vascular resistance. In extreme hemodilution blood viscosity is too low and insufficient to maintain functional capillary density, a problem that in experimental studies is shown to be corrected by increasing plasma viscosity up to 2.2 cP. This effect is mediated by Nitric oxide (NO) production via restoration of shear stress at the endothelium as shown by microelectrode perivascular measurements of NO concentration. Moderate elevations of blood viscosity by increasing hematocrit (approximately 10% of baseline) result in reductions of blood pressure by 10 mmHg of baseline. This effect is also NO mediated since it is absent after N-nitro-L-arginine methyl ester (L-NAME) treatment and in endothelial NO synthase deficient mice. These results show that the rheological properties of plasma affect vessel diameter in the microcirculation leading to counterintuitive responses to the increase in viscosity.

Expression of the antiangiogenic factor 16K hPRL in human HCT116 colon cancer cells inhibits tumor growth in Rag1(-/-) mice.

The M(r) 16,000 NH(2)-terminal fragment of human prolactin (16K hPRL) is a potent antiangiogenic factor inhibiting endothelial cell function in vitro and neovascularization in vivo. The present study was undertaken to test the ability of 16K hPRL to inhibit the growth of human HCT116 colon cancer cells transplanted s.c. into Rag1(-/-) mice. For this purpose, HCT116 cells were stably transfected with an expression vector encoding a peptide that included the signal peptide and first 139 amino acid residues of human prolactin (HCT116(16K)). Stable clones of HCT116(16K) cells secreted large amounts of biologically active 16K hPRL into the culture medium. Growth of HCT116(16K) cells in vitro was not different from wild-type HCT116 (HCT116(wt)) or vector-transfected HCT116 (HCT116(vector)) cells. Addition of recombinant 16K hPRL had no effect on the proliferation of HCT116(wt) cells in vitro. Tumor growth of HCT116(16K) cells implanted into Rag1(-/-) mice was inhibited 63% in four separate experiments compared with tumors formed from HCT116(wt) or HCT116(vector) cells. Inhibition of tumor growth of HCT116(16K) cells was correlated with a decrease in microvascular density by 44%. These data demonstrate that biologically active 16K hPRL can be expressed and secreted from human colon cancer cells using a gene transfer approach and that production of 16K hPRL by these cells was capable of inhibiting tumor growth and neovascularization. These findings support the potential of 16K hPRL as a therapeutic agent for the treatment of colorectal cancer.

16K human prolactin inhibits vascular endothelial growth factor-induced activation of Ras in capillary endothelial cells.

Signaling pathways mediating the antiangiogenic action of 16K human (h)PRL include inhibition of vascular endothelial growth factor (VEGF)-induced activation of the mitogen-activated protein kinases (MAPK). To determine at which step 16K hPRL acts to inhibit VEGF-induced MAPK activation, we assessed more proximal events in the signaling cascade. 16K hPRL treatment blocked VEGF-induced Raf-1 activation as well as its translocation to the plasma membrane. 16K hPRL indirectly increased cAMP levels; however, the blockade of Raf-1 activation was not dependent on the stimulation of cAMP-dependent protein kinase (PKA), but rather on the inhibition of the GTP-bound Ras. The VEGF-induced tyrosine phosphorylation of the VEGF receptor, Flk-1, and its association with the Shc/Grb2/Ras-GAP (guanosine triphosphatase-activating protein) complex were unaffected by 16K hPRL treatment. In contrast, 16K hPRL prevented the VEGF-induced phosphorylation and dissociation of Sos from Grb2 at 5 min, consistent with inhibition by 16K hPRL of the MEK/MAPK feedback on Sos. The inhibition of Ras activation was paralleled by the increased phosphorylation of 120 kDa proteins comigrating with Ras-GAP. Taken together, these findings show that 16K hPRL inhibits the VEGF-induced Ras activation; this antagonism represents a novel and potentially important mechanism for the control of angiogenesis.

The antiangiogenic factor 16K PRL induces programmed cell death in endothelial cells by caspase activation.

We asked whether the antiangiogenic action of 16K human PRL (hPRL), in addition to blocking mitogen-induced vascular endothelial cell proliferation, involved activation of programmed cell death. Treatment with recombinant 16K hPRL increased DNA fragmentation in cultured bovine brain capillary endothelial (BBE) and human umbilical vein endothelial (HUVE) cells in a time- and dose-dependent fashion, independent of the serum concentration. The activation of apoptosis by 16K hPRL was specific for endothelial cells, and the activity of the peptide could be inhibited by heat denaturation, trypsin digestion, and immunoneutralization, but not by treatment with the endotoxin blocker, polymyxin-B. 16K hPRL-induced apoptosis was correlated with the rapid activation of caspases 1 and 3 and was blocked by pharmacological inhibition of caspase activity. Caspase activation was followed by inactivation of two caspase substrates, poly(ADP-ribose) polymerase (PARP) and the inhibitor of caspase-activated deoxyribonuclease (DNase) (ICAD). Furthermore, 16K hPRL increased the conversion of Bcl-X to its proapoptotic form, suggesting that the Bcl-2 protein family may also be involved in 16K hPRL-induced apoptosis. These findings support the hypothesis that the antiangiogenic action of 16K hPRL includes the activation of programmed cell death of vascular endothelial cells.