Brachymesophalangia-5 without cone-epiphysis mid-5 in down's syndrome

Brachymesophalangia-5 proved to be far more frequent in 212 cases of Down's syndrome karyotype (i.e., 21%) than in 14,197 survey volunteers of European ancestry (1.4%). However, none of 28 juvenile Down's syndrome patients with brachymesophalangia-5 exhibited a cone-epiphysis on mid-5, as against the 47% that would be expected. Apparently the manifestation of brachymesophalangia-5 in the 47,G + karyotype is not simply a dosage effect associated with trisomy of chromosome 21.     

Prenatal origins of brachymesophalangia-5

As shown in 91 embryos and fetuses in the 41–235 mm crown-rump length, brachymesophalangia-5 may be identified in prenatal time. Although the ratios involving different combinations of length of mid-5 relative to other segments yield different frequencies of BMP-5, the short middle segment of the little finger is already associated with clinodactyly and developmental eccentricity as early as the ninth week of prenatal development.     

Cone-shaped epiphyses in Japanese children

Cone-shaped epiphyses (CSE) are reported in left-hand radiographs of a sample of 1,399 otherwise normal Japanese children aged 5–11 years. CSE occurred in only three centers: distal thumb, mid-index finger, and mid-fifth finger. The overall frequency of 23% shows a female bias (32% to 16%), which disappears when mid-5 cones are excluded. Chi-squared analyses show significant gender bias for mid-5 cones alone or in combination with the other phalanges, and show significant center associations: CSE in distal-1 and/or mid-2 are more common in association with mid-5 cones than they are without mid-5 cones. The CSE female bias in mid-5 is essentially unchanged across age-cohorts when bone-age is controlled for precocity, and no significant trends in CSE frequency with age are noted for either gender.     

Size reduction associated with brachymesophalangia-5: a possible selective advantage

The skin reflectance characteristics of a group of Quechua Indians have been described with an emphasis upon the effects of varying degrees of hybridization, sex and age. This group of Peruvian Indians occupied a reflectance range common to that of all other reported groups of South American Indians. Miscegenation with European Whites had a statistically significant although small influence upon skin color. In general males were consistently darker than females on the three body sites measured. A significant darkening on unexposed body areas occurred in both sexes during early adolescence which may have been caused by the high activity level of the pituitary gland at that stage of the growth cycle.     

Digenic Inheritance of Early-Onset Glaucoma: CYP1B1, a Potential Modifier Gene

"Early-onset glaucoma" refers to genetically heterogeneous conditions for which glaucoma manifests at age 5-40 years and for which only a small subset is molecularly characterized. We studied the role of MYOC, CYP1B1, and PITX2 in a population (n=60) affected with juvenile or early-onset glaucoma from the greater Toronto area. By a combination of single-strand conformation polymorphism and direct cycle sequencing, MYOC mutations were detected in 8 (13.3%) of the 60 individuals, CYP1B1 mutations were detected in 3 (5%) of the 60 individuals, and no PITX2 mutations were detected. The range of phenotypic expression associated with MYOC and CYP1B1 mutations was greater than expected. MYOC mutations included cases of juvenile glaucoma with or without pigmentary glaucoma and mixed-mechanism glaucoma. CYP1B1 mutations involved cases of juvenile open-angle glaucoma, as well as cases of congenital glaucoma. The study of a family with autosomal dominant glaucoma showed the segregation of both MYOC and CYP1B1 mutations with disease; however, in this family, the mean age at onset of carriers of the MYOC mutation alone was 51 years (range 48-64 years), whereas carriers of both the MYOC and CYP1B1 mutations had an average age at onset of 27 years (range 23-38 years) (P=.001). This work emphasizes the genetic heterogeneity of juvenile glaucoma and suggests, for the first time, that (1) congenital glaucoma and juvenile glaucoma are allelic variants and (2) the spectrum of expression of MYOC and CYP1B1 mutations is greater than expected. We also propose that CYP1B1 may act as a modifier of MYOC expression and that these two genes may interact through a common pathway.     

Lack of repeat transduction by recombinant adeno-associated virus type 5/5 vectors in the mouse airway

While recombinant adeno-associated virus (rAAV) vectors promote long-term transgene expression in the lungs and other organs, the goal of correcting chronic inherited lung diseases such as cystic fibrosis with this type of viral gene transfer vector is limited by the requirement of achieving stable potent transgene expression, potentially requiring vector readministration. Here we evaluated the abilities of rAAV type 5/5 (rAAV5/5) vectors based on the genome and capsid of AAV5 to efficiently transduce the lungs and nasal epithelium of mice after repeated administration. Transduction efficiency as judged by reporter gene expression was markedly reduced on a second rAAV5/5 administration and effectively abolished on a third. Varying the period between administrations from 8 to 36 weeks did not allow efficient repeated administration. A rapid rise in anti-AAV5 antibodies was noted after rAAV5/5 vector administration that was sustained for the entire period of investigation (in some cases exceeding 9 months). Furthermore, this antibody response and subsequent failure to repeatedly administer the vector were not rescued by the in vivo expression of CTLA4Ig from an rAAV5/5 vector. These results suggest that without the development of an effective and clinically acceptable immunosuppression strategy, treatments for chronic diseases that require repeated administration of rAAV5/5 vectors will be unsuccessful.     

Tissue-specific regulation of juvenile hormone esterase gene expression by 20-hydroxyecdysone and juvenile hormone in Bombyx mori

Juvenile hormone esterase (JHE) is the primary juvenile hormone (JH) metabolic enzyme in insects and plays important roles in the regulation of molt and metamorphosis. We investigated its mRNA expression profiles and hormonal control in Bombyx mori larvae. JHE mRNA was expressed at the end of the 4th and 5th (last) larval instars in the midgut and in all the three (anterior, middle, posterior) parts of the silk gland. In the fat body, JHE expression peaked twice in the 5th instar, at wandering and before pupation, while it gradually decreased through the 4th instar. When 20-hydroxyecdysone (20E) was injected into mid-5th instar larvae, JHE mRNA expression was induced in the anterior silk gland but suppressed in the fat body. Topical application of a juvenile hormone analog fenoxycarb to early-5th instar larvae induced JHE expression in both tissues. In the anterior silk gland, JHE expression was accelerated and strengthened by 20E plus fenoxycarb treatments compared with 20E or fenoxycarb single treatment, indicating positive interaction of 20E and JH. JHE mRNA is thus expressed in tissue-specific manners under the control of ecdysteroids and JH.     

Temperature and the Growth of Plant Cells

In cell elongation, the juvenile cell vacuolates, takes up water, and expands by irreversible extension of the growth-limiting primary walls. This process was elaborated analytically by Lockhart in the mid-1960s. His growth equation does not, however, include the influence of the environmental temperature at which cell growth takes place. In this article we consider a phenomenological model including temperature in the equation of growth. Also, by introducing the possible influence of growth regulators treated here as external perturbations, linear and nonlinear solutions are found. A comparison of experimental and theoretical results permits qualitative and quantitative conclusions concerning change in the magnitude of the cell wall yielding coefficient Φ as a function of both time and temperature (with or without external perturbations), which has acquired reasonable values throughout.     

Rank relations of juvenile and subadult natal males of Barbary macaques (Macaca sylvanus) at Affenberg Salem

Rank relations of more than 100 juvenile and subadult natal Barbary macaque males were analyzed. Hierarchical relations among individuals of the same age were established early during the first year of life. With few exceptions concerning infants from very high-ranking genealogies, males dominated female peers regardless of maternal rank. Males started to outrank females from older cohorts during the second year of life and completed the process of rank reversal with adult females at 5-6 years of age. An age-graded dominance pattern existed among males from different birth cohorts. Only 3 rank reversals between males from different cohorts were observed. Rank reversals among males of the same birth cohort occurred more frequently. Rank position of a male among his male peers was influenced by birth order, by maternal rank, and by the presence of juvenile brothers. Most males without juvenile brothers had low positions, regardless of maternal rank. Males born late in the birth season were also low-ranking, even when juvenile brothers were present. There was no cohort where ranking among males was determined by maternal rank alone, as is the case in rhesus monkeys and Japanese macaques. Adult/subadult male carriers had no noticeable effect on rank positions of 'their' infants. It is suggested that a weaker influence of Barbary macaque mothers on rank of their sons is related to very early integration of male infants in male social/play groups.     

Effects of photoperiod and social variables on reproduction and growth in the male musk shrew (Suncus murinus)

In Exp. 1, the time course for the photoperiodic response in juvenile male musk shrews was examined by exposing animals to short (10L:14D) or long (14L:10D or 18L:6D) daylengths for 10, 20, 40 or 56 days. When compared with the response of animals maintained in long days, those exposed to short days showed an inhibition of reproductive maturation by 40 days of treatment. In Exp. 2, the combined effects of photoperiod and social cues were investigated in juvenile males that were either housed in short days (with or without a female) or in long days (with or without a female) for 40 days. The short photoperiod was generally inhibitory to sexual maturation, while the presence of an adult female was generally stimulatory, i.e. animals living with a female in long days had the greatest level of sexual maturity, while those living alone in short days had the lowest level of maturity. Animals that received opposing signals, i.e. short days in the presence of a female, had an intermediate response which was equivalent to the response seen in males living alone in long days. These results suggest that the presence of a female can partly reverse inhibitory effects of short days. In Exp. 3, the effects of photoperiod and social cues on the reproductive physiology of the adult male (same design as in Exp. 2) were investigated. The results show that the adult male musk shrew is responsive to both photoperiodic and social cues, in a manner similar to that of juvenile animals.(ABSTRACT TRUNCATED AT 250 WORDS)     

Therapeutic Effects of Stem Cells and Substrate Reduction in Juvenile Sandhoff Mice

Sandhoff Disease (SD) involves the CNS accumulation of ganglioside GM2 and asialo-GM2 (GA2) due to inherited defects in the β-subunit gene of β-hexosaminidase A and B (Hexb gene). Substrate reduction therapy, utilizing imino sugar N-butyldeoxygalactonojirimycin (NB-DGJ), reduces ganglioside biosynthesis and levels of stored GM2 in SD mice. Intracranial transplantation of Neural Stem Cells (NSCs) can provide enzymatic cross correction, to help reduce ganglioside storage and extend life. Here we tested the effect of NSCs and NB-DGJ, alone and together, on brain β-hexosaminidase activity, GM2, and GA2 content in juvenile SD mice. The SD mice received either cerebral NSC transplantation at post-natal day 0 (p-0), intraperitoneal injection of NB-DGJ (500 mg/kg/day) from p-9 to p-15, or received dual treatments. The brains were analyzed at p-15. β-galactosidase staining confirmed engraftment of lacZ-expressing NSCs in the cerebral cortex. Compared to untreated and sham-treated SD controls, NSC treatment alone provided a slight increase in Hex activity and significantly decreased GA2 content. However, NSCs had no effect on GM2 content when analyzed at p-15. NB-DGJ alone had no effect on Hex activity, but significantly reduced GM2 and GA2 content. Hex activity was slightly elevated in the NSC + drug-treated mice. GM2 and GA2 content in the dual treated mice were similar to that of the NB-DGJ treated mice. These data indicate that NB-DGJ alone was more effective in targeting storage in juvenile SD mice than were NSCs alone. No additive or synergistic effect between NSC and drug was found in these juvenile SD mice.     

ENHANCEMENT OF 5-FLUOROURACIL CYTOTOXICITY BY CISPLATIN IN BRAIN TUMOUR CELL LINES

Cisplatin reportedly plays an important role as a chemical modulator in enhancing the chemotherapeutic effects of 5-fluorouracil on tumour cells. The aim of the present study was to test the synergistic cytotoxicity of cisplatin and 5-fluorouracil in 5-fluorouracil-resistant (C6) and -sensitive (9L) rat brain tumour cell lines. Survival fractions, determined using colony-formation assays, were compared following 5-fluorouracil treatment, with and without cisplatin. The presence of cisplatin (1–10μm ) enhanced cytotoxicity by more than three times compared with 5-fluorouracil alone in 5-fluorouracil-resistant C6 cells, whereas no enhancement effects were noted in 9L cells. These results suggest that a cisplatin-fluorouracil-based regimen may be promising in the treatment of 5-fluorouracil-resistant brain tumours.     

Quantitative MUC5AC and MUC6 mucin estimations in gastric mucus by a least-squares minimization method

We have determined the molar proportions of the MUC5AC and MUC6 mucus glycoproteins (mucins) in mucus from the normal and pathological human gastric antrum using a least-squares minimization analysis applied to amino acid compositions. We noted that the content of MUC5AC mucin in mucus from individuals without gastroduodenal disease was very high, suggesting that the integrity and barrier properties of the adherent gastric mucus layer are normally maintained by building-block structures formed from this mucin alone. We observed that the molar content of MUC6 mucin doubled (without significance) in mucus from patients with duodenal ulcer, and increased five times (with high significance) in mucus from patients with gastric ulcer, when compared with that in mucus from individuals without gastroduodenal disease.     

Synergistic effects of Actinobacillus suis cells in combination with 5-fluorouracil on mice bearing murine L1210 leukemia cells

 The mean survival rates of female BDF₁ mice transplanted intravenously (i.v.) with murine L1210 leukemia cells were significantly prolonged by intraperitoneal (i.p.) pretreatment (before i.v. transplantation) or by i.p. pre- and post-treatment (before and after the i.v. transplantation) with heat-killed Actinobacillus suis cells ATCC 15557 (AS 15557) alone, as compared with untreated L1210-leukemia-cell-bearing control mice. However, significant prolongation of the mean survival rates was not elicited by the i.p. post-treatment with AS 15557 alone. When 5-fluorouracil (5-FU) was applied i.p. to mice receiving post-treatment with AS 15557 alone, the mean survival rates of the L1210-leukemia-cell-bearing mice were significantly prolonged. The antileukemic action of AS 15557, alone or in combination with 5-FU, against L1210 leukemia was superior to that of a streptococcal preparation (OK-432) and was almost the same as of bacillus Calmette-Guérin with or without 5-FU. The results suggest the possibility that the synergism of AS 15557 in combination with 5-FU may be dependent on the relationship between the indirect immunological function of AS 15557 and the direct cytotoxic action of 5-FU on L1210 leukemia cells. Received: 5 December 1996 / Accepted: 28 March 1997     

Calcium ionophore enables soluble agonists to stimulate macrophage 5-lipoxygenase

The regulation of arachidonic acid conversion by the 5-lipoxygenase and the cyclooxygenase pathways in mouse peritoneal macrophages has been studied using particulate and soluble agonists. Particulate agonists, zymosan and latex, stimulated the production of cyclooxygenase metabolites as well as the 5-lipoxygenase product, leukotriene C4. In contrast, incubation with the soluble agonist phorbol myristate acetate or exogenous arachidonic acid led to the production of cyclooxygenase metabolites but not leukotriene C4. We tested the hypothesis that the 5-lipoxygenase, unlike the cyclooxygenase, requires activation by calcium before arachidonic acid can be utilized as a substrate. Addition of phorbol myristate acetate to macrophages in the presence of calcium ionophore (A23187) at a concentration which alone did not stimulate arachidonate metabolism resulted in a synergistic increase (50-fold) in leukotriene C4 synthesis compared to phorbol ester or A23187 alone. No such effect on the cyclooxygenase pathway metabolism was observed. Exogenous arachidonic acid in the presence of A23187 produced similar results yielding a 10-fold greater synthesis of leukotriene C4 over either substance alone without any effects on the cyclooxygenase metabolites. Presumably, calcium ionophore unmasked the synthesis of leukotriene C4 from phorbol myristate acetate-released and exogenous arachidonate by elevating intracellular calcium levels enough for 5-lipoxygenase activation. These data indicate that once arachidonic acid is released from phospholipid by an agonist, it is available for conversion by both enzymatic pathways. However, leukotriene synthesis may not occur unless intracellular calcium levels are elevated either by phagocytosis of particulate agonists or with calcium ionophore.     

Molecular screening of the major mutations in the ARSA gene in patients with metachromatic leukodystrophy

Metachromatic leukodystrophy (MLD) is an inherited storage disease caused by deficiency of arylsulfatase A (ARSA). Molecular analysis of the major mutations in the ARSA gene was performed in 10 Ukrainian patients (from 9 families) with MLD. According to the age of onset, late infantile MLD was identified in 3 patients, juvenile MLD in 5 patients, and adult MLD in 2 patients (sibs), respectively. The ARSA activity in the patients was 2-26 nmol/h/mg protein (the normal activity has been established in our laboratory as 111.9 +/- 7.1 nmol/h/mg protein). No correlation between enzyme activity and a clinical course of disease was revealed. The IVS2 + 1 mutation was found at 2 of 20 alleles (in a patient with late infantile form) and the P426L mutation was found at 2 of 20 alleles (in two patients with juvenile form). Thus, the total frequency of these two major mutations in the ARSA gene is 20% in Ukrainian MLD patients.     

Early changes in retinal 5'-nucleotidase activity in hereditary retinal degeneration

Activity of soluble cGMP phosphodiesterase (PDE) and of two membrane enzymes, 5'-nucleotidase and Na,K-ATPase, was studied in the developing retina of rats with inherited retinal degeneration. It was found that by day 10 of life, the content of 5'-nucleotidase in the afflicted rats was significantly reduced as compared with controls. This difference was unchanged throughout the subsequent animals' life. Na,K-ATPase activity in the afflicted and normal animals was the same. Within the first 45 days of life, PDE calculated with respect to the rhodopsin content was not different as regards both the afflicted and normal rats. When calculated with respect to protein, the changes in PDE corresponded with the reported data. The data obtained allowed a suggestion to be made that changes in 5'-nucleotidase in inherited retinal degeneration are disease-specific. They are accounted for by changes in the enzymes of nonphotoreceptor retinal membranes. The changes in PDE may be regarded as secondary, correlating with variation in the number of the photoreceptor membranes.     

Local habitat-induced variations in the population dynamics of Patella vulgata L.

The recruitment, growth and maximum size, life span, standing crop and biomass of Patella vulgata L. reveal considerable local differences in a range of habitats on the same shore. While some differences are related to physical conditions (e.g., standing water, tidal level), others arise from local biological interactions and may reverse expectations based on tidal level alone. Wet and bare surfaces apart, juvenile input is regularly highest among mid- and upper-shore mussels, while growth and maximum size are least among barnacles at any tidal level. The extreme permutations of population characteristics range from high-level, dry barnacle areas where limpet recruitment averages 15 % of standing crop, growth rate and maximum size are low and survival for 15–17 years is possible to low-level, bare rock where input averages 66 %, growth and maximum size are high and life span unlikely to exceed 4–5 years. Population densities vary annually but the relative densities of the different habitats have remained fairly steady as long as the biological conditions have remained similar; the latter largely control the recruitment and survival characteristics upon the balance of which the population density appears to depend. Although liable to distortions due to years of abnormal recruitment, the size-frequency structure of their populations is highly diagnostic of many habitats.     

Effects of a non-IGF binding mutant of IGFBP-5 on cell death in human breast cancer cells

We have demonstrated previously that IGFBP-5 alone had no effect on cell death but modulated ceramide-induced apoptosis in Hs578T IGF non-responsive cells. To investigate if IGFBP-5 maintains its intrinsic ability to modulate apoptosis in IGF-responsive cells, we used a non-IGF binding mutant of IGFBP-5. In Hs578T cells, non-glycosylated, glycosylated or mutant IGFBP-5 alone each had no effect on cell death, whereas all forms inhibited ceramide-induced apoptosis. In IGF-responsive MCF-7 cells, each wild type form reduced ceramide-induced cell death but mutant IGFBP-5 was without effect. In the presence of mutant IGFBP-5, however, IGF-I no longer conferred survival and in the presence of wild type IGFBP-5, long R3 IGF-I was also unable to confer survival. In summary, all forms of IGFBP-5 modulated ceramide-induced apoptosis in Hs578T cells. In MCF-7 cells, IGF-I-induced survival could be facilitated by IGFBP-5, but also blocked by IGFBP-5 if association with IGFBP-5 was prevented.