Necrotizing enterocolitis after intravitreal bevacizumab in an infant with Incontinentia Pigmenti - a case report.

BACKGROUND: Incontinentia Pigmenti is a rare disease affecting multiple organs. Fifty of patients show affection of the eye with retinopathy and possible amaurosis being the worst outcome. Treatment has commonly been panretinal laser coagulation but intravitreal application of bevacizumab as VEGF-inhibitor has shown to effectively suppress retinal neovascularization. CASE PRESENTATION: A six-week-old female infant with Incontinentia Pigmenti developed a foudroyant necrotizing enterocolitis shortly after intravitreal injection of bevazicumab due to a retinopathy with impending tractional detachment of the left eye. Since the onset of abdominal symptoms occurred immediately after the intravitreal application, a link between the two events seemed likely. Sequential analyses of the VEGF serum concentrations showed a massive suppression of endogenous VEGF with only a very slow recovery over weeks. Such a severe systemic adverse event has not been reported after intravitreal treatment with bevacizumab in an infant. CONCLUSION: This case report shows a relevant systemic uptake of bevacizumab after intravitreal application as suppressed VEGF levels show. There seems to be a connection between suppressed VEGF levels and the onset of necrotizing enterocolitis. Therefore, treatment with bevacizumab should be carefully considered and further research is needed to assess this drug's safety profile.

Is there a secular decline in motor skills in preschool children?

Current research seems to confirm a secular decline in movement skills in school children. Only few data are available for preschool children and no clear trend can be identified. In the year 2007, height, weight, and motor performance were determined in 726 preschool children [Prevention through Activity in Kindergarten Trial (PAKT)] and compared with historical samples from 1973, 1985, and 1989. There was no difference in height and weight between the samples of 1973 and 2007. Older boys of today were smaller and lighter than those of 1989. Regardless of age, PAKT children fared significantly better in standing long jump than those assessed in 1989. Compared with the sample of 1973, PAKT children did equally well in this task. There were no differences in performance in an obstacle course between children of 1989 and 2007. In balancing backwards, PAKT children performed significantly worse than those in 1985. Regarding target throwing only the PAKT 4-year-olds achieved significantly worse results than those in 1985.Therefore, in preschool children, a secular decline is only evident in some, but not all, motor skills, which may indicate a change in behavior activity over the last decades.

Synthesis and properties of 5,6-dichlorobenzimidazole 2'----5'- and 3'----5'-nucleotide dimers and trimers.

5,6-Dichloro-1-beta-D-ribofuranosylbenzimidazole (2), synthesised by the fusion method, was used for the synthesis of 2'----5'- and 3'----5'-linked di- and tri-meric oligonucleotides. The protecting groups used were p-methoxytrityl for HO-5', tert-butyldimethylsilyl for HO-2',3', and 2,5-dichlorophenyl and 2-(4-nitrophenyl)ethyl for the phosphate group. The internucleotidic linkages were established by the phosphotriester approach to give the fully protected 2'----5' dimers (15, 17, and 18) and trimers (27 and 28), as well as the 3'----5' dimers (22 and 23) and trimers (31 and 32). Deprotection involved a sequence of steps to afford the corresponding free oligonucleotides 21, 26, 30, and 33 isolated as the triethylammonium salts in good yields. The new compounds were characterised by elemental analysis and by u.v. and 1H-n.m.r. spectroscopy.

Changes in the dynamics of luteinizing hormone-releasing hormone-stimulated secretion of luteinizing hormone during sexual maturation of female rats.

Our aim was to identify age-related changes in the dynamics of luteinizing hormone (LH) release that may contribute to the decline in pituitary sensitivity to luteinizing hormone-releasing hormone (LHRH) during sexual maturation of female rats. We studied LHRH-stimulated LH secretion curves of superfused pituitaries from rats ranging in age from 10 days to the first estrous cycle. Pituitary fragments were exposed for 10 min to medium alone or to medium plus LHRH; incubation continued in medium alone for 130 min and effluent was collected for LH analysis. Secretion curves were compared on the basis of total secretion (area under the curve), maximal change in LH secretion rate, and rates of rise and decay of the curves. The data show that total LH secretion in response to LHRH is greatest in 15-, 20-day-old and first-proestrus animals. Also, the maximal change in LH secretion rate was greater, and the increase in LH secretion rate faster in younger animals than in 30-day-old animals. Analysis of secretory granules in LH-containing gonadotropes of 15- and 30-day-old animals revealed changes in he granule population with age. We conclude that younger animals respond faster with a greater LH secretion response to LHRH than do 30-day-old or first-estrus animals, and that these age-related changes in the dynamics of LH secretion may be due in part to maturation of the LH secretory granules.

Plasma concentrations of free 5 alpha-androstane-3 alpha, 17 beta-diol and related gonadal steroids during spontaneous and induced sexual maturation in the female rat.

Plasma concentrations of free 5 alpha-androstane-3 alpha, 17 beta-diol (3 alpha-diol), 5 alpha-androstane-3 beta, 17 beta-diol (3 beta-diol), 5 alpha-dihydrotestosterone (DHT) and of testosterone (T) were determined in female rats undergoing spontaneous sexual maturation or pregnant mare's serum gonadotropin (PMSG)-induced precocious ovulation. In the prepubertal period, 3 alpha-diol levels showed an inverse relationship with uterine weights and a significant drop between Days 21 and 26 of life. Postpubertally, 3 alpha-diol concentrations tended to rise again, to a lesser degree, in parallel with increments in plasma T. Concentrations of 3 alpha-diol were increased tenfold 48 h after the administration of PMSG, i.e., at a time when T and estradiol-17 beta also peaked. Concentrations of 3 beta-diol and DHT remained low throughout both experiments. In earlier studies, it was found that exogenous 3 alpha-diol suppresses reproductive function in the immature rat, but the high concentrations of endogenous 3 alpha-diol reported here for the PMSG model clearly do not interfere with ovulation. The observed constellations of plasma steroids are compatible with the postulate of a peripubertal shift in ovarian progesterone metabolism from 5 alpha-reduced C21 steroids to delta 4-3-ketosteroids.

On the significance of 5 alpha-androstane-3 alpha,17 beta-diol in the peripubertal female rat.

The biological effects of exogenous 5 alpha-androstane-3 alpha,17 beta-diol (3 alpha-diol) and its 3 beta-epimer (3 beta-diol) on sexual maturation were assessed in Sprague-Dawley rats. Chronic administration of 3 alpha-diol, but not of 3 beta-diol, prevented ovulation and suppressed uterine and ovarian development. Vaginal opening was advanced by 3 alpha-diol, but neither vaginal cytology nor ovarian histology revealed any signs of ovarian cyclicity. When administered acutely, 3 alpha-diol prevented the estrogen-priming required for the triggering of gonadotropin surges by progesterone as well as the estrogen-sensitization of pituitary gonadotrophs to LHRH stimulation. Plasma concentrations of endogenous free 3 alpha- and 3 beta-diol and of testosterone and dihydrotestosterone were assessed during spontaneous and PMSG-induced sexual maturation. The observations are compatible with the postulate of a puberty-related shift in ovarian biosynthetic pathways from the preferential 5 alpha-reduction of progesterone to the production of aromatizable androgens.

Effect of catechol oestrogens on induced ovulation in the immature rat.

The 'positive feedback' effect of exogenous oestradiol-17 beta in advancing ovulation induced by pregnant mare serum gonadotrophin (PMSG) has been used in the present study as a model in which to test the possible oestrogenic or antioestrogenic effects of the catechol oestrogens, 2-hydroxyoestradiol (2-OHE2) and 4-OHE2. Sprague-Dawley rats of 26 days of age were injected with 20 i.u. PMSG together with either vehicle alone or test steroids. The animals were killed 72 h later and the Fallopian tubes were examined for the presence of ova. Advancement of induced ovulation by treatment with oestradiol was confirmed; 2-OHE2, in doses of up to 100 micrograms, influenced neither the time of ovulation nor the number of ova present but 4-OHE2 was equipotent with oestradiol in doses varying from 0.5 micrograms (the minimum effective dose for both steroids) to 10 micrograms. The possible antioestrogenic effect of 2-OHE2 was tested by giving a 100 micrograms dose either at the same time or 2 h before PMSG plus 2 micrograms oestradiol or 4-OHE2. The effects of oestradiol and 4-OHE2 were not altered by this treatment. These data show that, in this model of 'positive feedback', 2-OHE2 has neither an oestrogenic nor an antioestrogenic action but that 4-OHE2 has a potent oestrogenic action, thus raising the question of a physiological role for 4-OHE2 in the regulation of ovulation.

Effects of estradiol-induced lesions of the arcuate nucleus on gonadotropin release in response to preoptic stimulation in the rat.

Female Wistar rats treated with a single subcutaneous injection of 2 mg estradiol valerate (EV) develop gradually progressive, multifocal lesions of the arcuate nucleus. They also exhibit vaginal estrus and endocrine profiles characteristic of animals sustaining anterior hypothalamic deafferentation. In this study, EV-treated females with the arcuate lesions released significantly less LH 1 h following electrochemical stimulation of the medial preoptic area (MPOA) than did normally cycling controls in proestrus. FSH release in response to MPOA stimulation was the same for both groups. As plasma LH concentrations were not significantly different between EV-treated and control animals 1 h after the injection of a potent LHRH analog, the reduced LH response to MPOA stimulation appears to reflect a primarily hypothalamic defect. However, the EV treatment also affected pituitary responsiveness to long-term stimulation as evidenced by reduced LH responses to the LHRH analog after 2 and 3 h. No such differences were seen in the FSH response.

Steroid induction of gonadotropin surges in the immature rat. I. Priming effects of androgens.

Sexually immature female rats were either primed with estradiol benzoate on day 23 or given daily injections of various androgens on days 23--25. Plasma for LH and FSH determinations was collected on day 26, 5 h after an injection of progesterone. Massive gonadotropin surges were found after priming with estradiol benzoate or treatment with dehydroepiandrosterone (DHEA), delta 4-androstenedione, and testosterone, but not with ring A-reduced androgens (5 alpha-dihydrotestosterone, 5 alpha-androstane-3 alpha,17 beta-diol, its 3 beta-epimer, and androsterone) or the nonaromatizable 11 beta-hydroxy- and 11-ketoderivatives of delta 4-androstenedione. Rats bearing DHEA-containing Silastic implants also produced LH surges in response to progesterone. A single injection of an antiestrogen antiserum abolished gonadotropin surges in rats primed with estradiol benzoate or DHEA and greatly reduced the accompanying uterine hypertrophy. DHEA and delta 4-androstenedione were barely uterotrophic in ovariectomized rats but sustained progesterone-induced gonadotropin surges. The results indicate that certain (adrenal?) androgens are able to induce maturation of the steroid-sensitive surge system via extragonadal aromatization, whereas their uterotrophic effect is largely mediated by the ovaries. Coordinated increased conversion of androgens at central and peripheral sites may be of physiological importance for the triggering of puberty.

Steroid induction of gonadotropin surges in the immature rat. II. Triggering ability of progesterone metabolites, adrenocortical hormones, and adrenocorticotropin.

Several adrenocortical steroids were tested for their ability to trigger LH release in estrogen-primed sexually immature female rats. Massive LH surges, approaching these known to be triggered by progesterone (P), followed the injection of depot ACTH1-24 and also of the adrenal steroids deoxycorticosterone (DOC) and 4-pregnen-21-ol-3,20-dione 21-acetate, but not of 5 alpha- and 5 beta-pregnan-21-of-3,20-dione, corticosterone, or aldosterone. Estrogen-primed adrenalectomized/ovariectomized rats also responded to DOC, albeit to a lesser extent. The P metabolites 5 alpha-pregnana-3,20-dione, 4-pregnen-20 alpha-ol-3-one, and 3 alpha-hydroxy-5 alpha-pregnan-20-one proved ineffective, although their triggering ability in adults was confirmed. It is concluded that adrenal P and DOC are potent activators of the gonadotropin surge system underlying pubertal ovulation and that P metabolites may acquire biological properties during sexual maturation.

The participation of luteinizing hormone releasing hormone (LRH) in the process of sexual maturation.

Precious pubertal ovulation and premature steroid-induced gonadotropin surges were triggered in 23-day old rats by unilateral electrolytic lesions placed in the basal hypothalamus. Attempts were made to reproduce both aspects of precocious sexual maturation by repeated injections or prolonged infusion of Luteinizing Hormone Releasing Hormone (LRH) or by repeated administration of the potent and long-acting analog (D-Leu6, des-Gly-NH2(10))-LRH-ethylamide. Neither effect of the brain lesion was reproduced by LRH or its analog. It is concluded that brain lesions advance puberty in this species by mechanisms not involving the discharge of LRH. The role of the adrenal cortex in providing estrogen precursors required for the induction of ovarian gonadotropin receptors is discussed.

In-vitro studies of the effects of oestrogen pretreatment on the sensitivity of the immature female rat pituitary gland to stimulation with gonadotrophin releasing hormone.

The effects of oestrogen priming on the sensitivity of the anterior pituitary gland to stimulation with gonadotrophin releasing hormone (GnRH) was investigated in immature female rats using a new organ culture technique. Hemipituitary glands obtained from animals primed with a single dose of oestradiol benzoate (OB; 20 microgram/100 g body weight) released significantly more LH when pulsed with GnRH (4 nmol/1) than did control hemipituitary glands. This potentiating effect was detectable as early as 5 days after birth. After a second stimulation, LH secretion remained high. These results were compared with those obtained from animals treated to induce increased levels of endogenous oestrogen on day 26 of life. Thus, hemipituitary glands were obtained from animals given two injections of OB, an injection of pregnant mare serum gonadotrophin (PMSG) or a unilateral brain lesion placed in the basal hypothalamus. Pituitary tissue was stimulated as before with a pulse of GnRH. Two injections of OB enhanced the sensitivity to stimulation. Conversely, both PMSG and lesion treatment severely reduced the sensitivity to GnRH, although PMSG-treated and lesioned animals have been used as models for the study of ovulation.

Anterior pituitary sensitivity in immature female rats stimulated with gonadotrophin releasing hormone in vivo: effect of priming with oestrogen, pregnant mare serum gonadotrophin or brain lesion.

PIP: Anterior pituitary sensitivity, assessed in terms of increments in plasma luteinizing hormone (LH) and follicle stimulating hormone (FSH) concentrations to stimulation with 1 or 2 injections of gonadotrophin releasing hormone (GnRH) was investigated in 26-day-old immature female rats which had received 1 of the following priming treatments: 1) 10 mcg estradiol benzoate (EB) as a single injection on Day 23 or Day 25 or on both days, 2) 10 IU pregnant mare serum gonadotrophin (PMSG) on Day 24, 3) an electrochemical brain lesion placed in the mediobasal hypothalamus on Day 23, or 4) control animals received either vehicle alone or a sham lesion. Pituitary sensitivity assessed at 1000 hours on Day 26 after 1 or 2 injections of GnRH was enhanced to a similar degree in the 3 groups treated with EB in terms of LH (p .01). FSH increased somewhat after EB treatment. In contrast, 48 hours after the injection of PMSG pituitary sensitivity, in terms of both LH and FSH, dropped sharply (p .001). Sensitivity to 1 injection of GnRH was unchanged in lesioned rats. However, a 2nd GnRH injection administered after an hour interval induced a slightly larger LH response in control animals. In another study, rats treated with EB on Day 23 and with 1 mg progesterone at 1200 hours on Day 26, pituitary sensitivity increased at both 1400 and 1700 hours as compared with that in the Day 23 EB-treated group at 1000 hours. PMSG-treated rats maintained their state of decreased responsiveness at 1400 hours, but exhibited increased sensitivity at the time of the gonadotropin surge (1700 hours).^ieng

Immediate effect of lesion of the ventromedial hypothalamic area upon glucose-induced insulin secretion in anaesthetized rats.

Insulin secretion, measured in vivo following an intravneous load of glucose to anaesthetized rats, was markedly increased ten minutes after bilateral electrolytic lesions of the ventromedial hypothalamic (VMH) area when compared to both sham-operated and unoperated controls. The successful lesioning of the VMH area was assessed by the subsequent occurrence of hyperphagia, as estimated by the increase in body weight. It is concluded that the ventromedial hypothalamic area exerts an inhibitory influence upon the secretory activity of the B-cells. Furthermore, the rapid disappearance of such inhibitory influence following lesions of the VMH suggests that this area of the brain may be of importance in the minute to minute regulation of insulin secretion. The precise anatomical location of the hypothalamic "nucleus" (or "nuclei") involved, as well as the neural or humoral nature of its inhibitory effect upon the endocrine pancreas remain to be elucidated.