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A pericentric inversion of an X chromosome [46,X,inv(X)(p11q28] with no detectable deletions was ascertained by amniocentesis for prenatal diagnosis in a 42-year-old woman. No defintive counsel could be offered as such an inversion had not been previously reported. At 1 year, the infant appears normal. These data and the review of other recently published karyotypes suggest an absence of a somatic position effect in the human X chromosome and possible biases of ascertainment or possible alternative karyotypic interpretations of some previously reported inv(X) cases.

Cyclically administered estrogen helps prevent and treat vasomotor symptoms and atrophic changes in the genitourinary tracts of postmenopausal women. Administering exogenous estrogen may also help prevent postmenopausal bone loss and help treat severe postmenopausal osteoporosis. Although recent studies have indicated an association between estrogen replacement therapy and endometrial carcinoma, a true cause-and-effect relationship has not been established. Thus, estrogen replacement therapy should be administered carefully, considering the risk/benefit ration for each individual, and should use the smallest dose necessary to achieve the desired effect. If irregular bleeding occurs before or during estrogen administration, a complete gynecologic examination (including a histologic examination of the endometrium) is mandatory.

Characterisation and pharmacological data of twelve synthetic tetrapeptide analogs of dermorphin (opioid heptapeptide) are reported. An N-terminal tetrapeptide free acid or ester is less potent that the corresponding amide. Whereas substitution of Gly4 by another aminoacid residue is well tolerated, substitution of D-Ala2 by D- or L-alpha-aminoxypropionic acid causes a complete loss of activity. Finally, guanidination of the tetrapeptides results in an increase of peripheral and central opioid activity.

Pharmacological activity in vitro and in vivo of 17 pentapeptide analogs of dermorphin (opioid heptapeptide) are reported. Substitutions of the Tyr5 residue afford dermorphin related pentapeptides which are likely to discriminate between mu- and delta-receptors.

The course of endomitosis in human hydatidiform moles has been analyzed. It differs from the classical description of endomitosis in that endoprophase is completely missing and, very probably, so is a typical interphase. The chromosomes are even less synchronized in their replication and condensation cycle than in normal mitosis. At no point do all chromosomes decondense, but a part remains condensed while others are extended and in the process of synthesizing DNA. Even two paired sister chromosomes may replicate nonsynchronously. The latest replicating chromosome is usually a large darkly staining chromosome, which we tentatively identify as the inactive X. No DNA synthesis takes place during "endometaphase" or "endoanaphase" stages, when the chromosomes are most condensed. Some polyploid "endoanaphases" or "endotelophases" with stretched out chromosomes obviously represent end-stages of the endomitotic pathway, and the nuclei are in the process of reverting into evenly stained nuclei. In some "endometaphases," a near-haploid number of chromosomes can be counted. In others, the endochromosomes seem to be compound structures consisting of several chromosomes that have not separated during the previous endomitoses. This is seen also in normal trophoblast and cervical cancer. In large cancer cells, such bundles can be seen in the process of falling into individual chromosomes.

The premeiotic and meiotic stages are described in the pollen mother cells of the liliaceous plant Eremurus. In human oocytes from embryonic ovaries, the premeiotic and early meiotic stages up to dictyotene have been identified on morphological grounds. In Eremurus, in which each stage can be independently verified by the sequence of buds situated in a spiral, there is no indication of somatic pairing of homologous chromosomes, nor is there any sign of a premeiotic contraction of the chromosomes. The interphase following this mitosis is, in turn, succeeded by leptotene in which the DNA synthesis occurs, as determined by using autoradiography. This stage is followed by a "distance pairing" stage in which the homologous chromosomes lie parallel to each other at a distance. In typical zygotene, segments of chromosomes are paired intimately, others are unpaired, and the points at which pairing begins and ends are clearly visible. Each bivalent shows several pairing blocks. Pairing is completed at pachytene; diplotene which is characterized by the separation of the chromosomes follows. In mid-diplotene the chromosomes collect together in the so-called second contraction stage. The same meiotic prophase stages that occur in Eremurus, including the "distance pairing" stage, are found in the embryonic human oocytes. In the last premeiotic interphase, the chromosomes appear as condensed prochromosomes which unravel directly to form the leptotene chromosomes. In the oocytes, too, DNA synthesis seems to take place in leptotene.

The concept and role of endomitosis is reevaluated in the light of observations on three organisms. Endomitosis which morphologically agrees with Geitler's (1939) classical definition is compared in tapetal cells of the liliaceous plant Eremurus, in the septal cells of the testicular follicles of the grasshopper Melanoplus, and in human cells from normal and molar trophoblasts and cervical cancer. These observations, together with those of Kidnadze and Istomina (1980), show that functionally at least two fundamentally different types of endomitosis exist, although morphologically the stages resemble each other in the three organisms. In the first type, exemplified by Eremurus, each endomitosis leads to a chromosome constitution which represents one level higher ploidy, a course that has been assumed to be characteristic of endomitosis in general. The second type, observed in its most characteristic form in the grasshopper, seems to be stationary: no DNA synthesis occurs, but an intensive RNA synthesis takes place. Presumably such cells have reached a final state in their development and are specialized in manufacturing one or more gene products. Endomitosis in normal placenta comes near this type, although DNA synthesis takes place in occasional cells. However, similar endomitotic nuclei in the hydatiform moles are in the process of DNA synthesis. When endomitosis is analyzed in different organisms and tissues, the observation that this process is not one entity should be kept in mind.

Eight new dermorphin tetrapeptides, X-Tyr-D-MetO-Phe-aa-Y (X = H, H2N = C(NH); aa = Gly, 2-aminoethanol, sarcosine; Y = NH2, NH-alkyl), were prepared and tested for opioid activity. They show dose-related naloxone-reversible opioid effects in vitro and in vivo. H-Tyr-D-MetO-Phe-Gly-NH2 (I) (guinea pig ileum IC50 = 13.6 nM; tail-flick ED50 = 1.97 pmol/mouse, icv, and 0.65 mumol/kg, sc), though less effective in the periphery, has central activities higher than those of dermorphin H-Tyr-D-Ala-Phe-Gly-Tyr-Pro-Ser-NH2. Following intracerebroventricular or subcutaneous administrations in mice, I is about respectively 1500 and 17 times as potent an analgesic as morphine.

H-Tyr-D-Ala-Phe-Gly-Tyr-Pro-Ser-NH2 (demorphin), and opiate-like peptide, and tri-, tetra-, penta- and hexapeptide-amide analogs, were synthesized by conventional methods in solution, to determine the minimum peptide chain-length, required for analgesic activity.

The correlations of abnormal X chromosome constitutions and the resulting phenotypes in the human female are reviewed. The following hypotheses put forward to explain these correlations are discussed in detail: (1) The damage is done before X inactivation; (2) An effect is exerted between reactivation of the X chromosome(s) and meiosis in oocytes; (3) A recessive gene(s) in hemizygous condition might be expressed in the cases in which the same X is active in all cells; (4) A change in the number of presumed active regions on the inactive X chromosomes might have an effect; (5) A position effect, in that the region Xq13-q27 has to be intact in both X chromosomes to allow normal development, may be responsible; (6) An effect during the period when cells with different inactivation patterns compete is a probability; (7) The original X inactivation may be neither regular nor random. The conclusion reached is that the phenotypic effects of a specific X chromosome aberration may be simultaneously exerted through different pathways (Tables 1 and 2). Hypotheses (2), (4), (5), and (6) are considered probable. Hypothesis (3) has been discarded, and there is very little evidence for hypotheses (1) and (7).

Segregation after mitotic crossing-over in an isodicentric (idic) X chromosome with one active and one inactive centromere has given rise to two new cell lines, one in which the idic(Xpter) chromosome has two active centromeres (most of these chromosomes also have an inversion) and another in which neither centromere is active. The two X chromosomes are attached at the telomeres of their short arms. Similar segregation has given rise to two other cell lines with idic(Xq-) chromosomes. Other observations on segregation after mitotic crossing-over are reviewed. Unequal crossing-over has apparently played a major role in the evolution of various genes and heterochromatin. Retinoblastoma and Wilms tumor are in some cases associated with homozygosity of a chromosome segment resulting from mitotic crossing-over. Similarly, the high incidence of cancer in Bloom syndrome may be caused by mitotic crossing-over leading to homozygosity or amplification of oncogenes.

The synthesis and pharmacological activity of Leu-enkephalins carrying a free or substituted carboxymethyl group at glycine-2-nitrogen are described.

The synthesis and in vitro pharmacological tests of 5 ketomethylene tetrapeptide analogues of the opioid heptapeptide dermorphin H-Tyr-D-Ala-Phe-Gly-Tyr-Pro-Ser-NH2 are described in this study. The substitution of the original Phe-Gly peptide bond (-CO-NH-) by the ketomethylene linkage (-CO-CH2-) provided analogues with reduced opioid activity.

A mentally retarded girl with several Turner symptoms had the chromosome constitution 46,Xtel(Xq). The abnormal X chromosome appeared to be completely telocentric and stable. It was late-replicating and formed a smaller than normal Barr body. The origin of telocentric chromosomes is discussed.

Preliminary pharmacological data and the characterization of sixteen synthetic tetrapeptide-amide analogs of dermorphin (opioid heptapeptide) are reported. N,N-dialkylamides displayed reduced or no activity; on the contrary, derivatization of H-Tyr-D-Ala-Phe-Gly-OH C-terminus by suitable amide moieties (1-adamantanamine, 1-adamantane-methylamino, or D-alpha-methylbenzylamine) produced potent analogues with peripheral and central opioid activities comparable to, or higher than those of dermorphins.

Cases of autosomal trisomy and trisomy mosaicism among liveborn infants are reviewed, and a second case of chromosome 3 trisomy mosaicism is described. The occurrence of autosomal trisomy for a particular chromosome is in general negatively correlated with the number of genes which have been localized to that chromosome. It is also positively related to the Q-brightness of the chromosome, which reflects its content of intercalary heterochromatin. Furthermore there are significantly fewer autosomal trisomics for chromosomes which contain hot spots for mitotic chiasmata in Bloom syndrome (chromosomes 1, 3, 6, 11, 12, 17, 19, and 22), compared with similar-sized control chromosomes 2, 4, 7, 9, 10, 18, 20, and 21. This is interpreted as further evidence for the gene richness of the hot spots which, being active, are extended in interphase and are therefore available for mitotic crossing over. The gene richness of these short Q-dark regions is also borne out by the scarcity of trisomic abortions for the chromosomes involved (the embryo dies before the abortion is recognized) and by the higher number of genes localized to these chromosomes compared with the control chromosomes.

The preliminary pharmacological activity pattern in guinea pig ileum and mouse vas deferens bioassays of five dermorphin related tetrapeptides is reported. The biological activities of the compounds under examination appear to be correlated in a statistically significant way to the lipophilic character of the C-terminal substituents.

The genetics of Müllerian aplasia (absent fallopian tubes, absent or rudimentary uterine corpus and cervix, absent upper vagina) has never been investigated systematically. Some investigators believe the disorder is inherited in female-limited autosomal dominant fashion, males transmitting the mutant gene but, of course, not manifesting the trait. To investigate this possibility, we obtained pedigrees in 23 probands with Müllerian aplasia. None had an affected relative. The absence of affected individuals among 30 postpubertal sisters, 31 paternal aunts, and 40 maternal aunts makes it unlikely that a sex-limited autosomal dominant gene is a common cause of Müllerian aplasia in our population. Dominant genes might exist in other populations, and fresh dominant mutations cannot be excluded. However, polygenic/multifactorial inheritance is perhaps more plausible.

The modification of the dermorphin-(1-4)-tetrapeptide structure led to analogues with potent opioid activity in vitro and in vivo. [Sar4]Tetrapeptides such as H2N-CH(NH)-Tyr-D-Ala-Phe-Sar-D-NH-CH(CH3)C6H5 (VII) whose terminal amino group is replaced by the guanidino function and whose C-terminus is amidated by (R)-(+)-alpha-methylbenzylamine, show peripheral and central opioid activities comparable to or higher than those of dermorphin. The potency of VII in the different tests was as follows: guinea-pig ileum (GPI) IC50 = 0.09nM, mouse Vas deferens (MVD) IC50 = 0.69nM, tail-flick ED50 = 8.91 pmol/mouse, i.c.v. and 4.54 mumol/kg, s.c. This dermorphin-(1-4)-tetrapeptide derivative is about 650 and 950 times as active as morphine in the two in vitro tests, respectively. The MVD/GPI potency ratio of the new peptides suggests a mu-type agonist behaviour.

The anti-inflammatory and analgesic activities of a series of 3-methyl-N-phenyl-1H-pyrazol-5-ylcarboxamides were investigated and compared with flufenamic acid. The compounds were synthesized by condensation of diketopiperazines 2 with the appropriate aniline. The pharmacological tests showed that some compounds have good anti-inflammatory activity in rat paw edema induced by carrageenin and low toxicity.

We describe the synthesis and preliminary in vitro and in vivo pharmacological tests of six [beta-Ala4]tetrapeptides analogs of the opioid heptapeptide dermorphin H-Tyr-D-Ala-Phe-Gly-Tyr-Pro-Ser-NH2. The modification obtained by substituting beta-Ala4 for Gly4 provided further analogues with significant opioid activity.

An idic(Xp--) in which the two X chromosomes are attached short arm to short arm, and which thus has two b regions (the Q-dark segment next to the centromere on Xp) between the inactivation centers, assumed to be situated on the Q-dark region next to the centromere on Xq, showed 63.8% bipartite Barr bodies as compared with 22.2% formed by idic(Xq--). In addition, the mean distance of the two parts of the Barr bodies in the fibroblasts of a patient with idic(Xp--) is significantly greater than in the cases with one or no b region. Contrary to the other patients with abnormal X chromosomes, the buccal cells of a woman idic(Xp--) showed a number of bipartite Barr bodies. -- To explain these observations we have put forward the hypothesis that the b region on the Xp always remains active and thus, when the rest of the chromosome forms a Barr body, this segment is extended, allowing the two parts of the X chromatin to get farther apart and at the same time increasing the percentage of bipartite bodies.

The interference of NA and 5-HT on Morphine inhibition on electrically-driven guinea-pig ileum was investigated. The alpha-antagonist Phentolamine (1 gamma/ml) and alpha MT-pretreatment reduced Morphine inhibition on muscle twitch. Similarly, ilea taken from pCPA-pretreated animals were less sensitive to the opioid. In contrast, both NA or 5-HT added either to normal or to amine-depleted tissue enhance Morphine inhibition. Therefore, not only in the brain but also in the guinea-pig ileum NA and 5-HT enhanced opioid effect.

The preliminary pharmacological data on synthetic peptides related to dermorphin (opiate-heptapeptide) are reported. The tetrapeptide, corresponding to the N-terminal sequence of dermorphin is a minimum requirement for significant opiate-activity in vitro.

A woman complaining of secondary amenorrhea and infertility was found to have, in addition to a normal X chromosome, a long compound chromosome that consisted of two X chromosomes attached by telomeric fusion of the short arms without any apparent loss of genetic material, 46,X,dic(X)(pter), hereafter referred to as t(X;X). The t(X;X) chromosome was late replicating; the inactivation pattern was analyzed. It formed Barr bodies that were larger than normal, of which the majority were bipartite. Analysis of the X-chromatin masses supports our hypotheses that there is a region next to the centromere on the short arm of the inactive X chromosome that remains active and that there is an inactivation center located on Xq, a short distance from the centromere, around which the X chromatin condenses.

Endopolyploidy, which arises through the duplication of DNA without accompanying nuclear division, occurs in large numbers of lower and higher plants and animals, including the best known, the salivary gland nuclei of Drosophila. Endomitosis is one of the processes leading to endopolyploidy, in which the stages of mitosis (prophase, metaphase, anaphase) take place inside the nuclear membrane and without spindle formation. In mammals, endomitosis has been observed in the trophoblast of the placenta of the mouse, rat and rabbit. This is the first report of endomitosis in a normal human tissue, the trophoblast of first trimester human placenta.

By employing the mouse tail-flick assay the analgesic activity of selected dermorphin tetrapeptides was assessed. The remarkable differences in potency exhibited by peptides after i.c.v. (500-1000 times higher than morphine) and s.c. (nearly comparable to morphine) administration are probably due to peptidase degradation.

The origin and behavior of human dicentric chromosomes are reviewed. Most dicentrics between two nonhomologous or two homologous chromosomes (isodicentrics), which are permanent members of a chromosome complement, probably originate from segregation of an adjacent quadriradial; such configurations are the result of a chromatid translocation between two nonhomologous chromosomes, or they represent an adjacent counterpart of a mitotic chiasma. The segregation of such a quadriradial may also give rise to a cell line monosomic for the chromosome concerned (e.g., a 45, X line). Contrary to the generally held opinion, isodicentrics rarely result from an isolocal break in two chromatids followed by rejoining of sister chromatids. In this case the daughter centromeres go to opposite poles in the next anaphase, and the resulting bridge breaks at a random point. This mechanism, therefore, leads to the formation of an isodicentric chromosome only if the two centromeres are close together, or if one centromere is immediately inactivated. Observations on the origin of dicentrics in Bloom syndrome support these conclusions. One centromere is permanently inactivated in most dicentric chromosomes, and even when the dicentric breaks into two chromosomes, the centromere is not reactivated. The appearance and behavior of the "acentric" X chromosomes show that their centromeres are similarly inactivated and not prematurely divided. Two Bloom syndrome lymphocytes, one with an extra chromosome 2 and the other with an extra chromosome 7, each having an inactivated centromere, show that this can also happen in monocentric autosomes.

The confluence of pseudo-pseudohypoparathyroidism, empty sella syndrome, and pituitary insufficiency is reported for the first time. The patient, an 18-year-old woman, was evaluated because of primary amenorrhea. Possible relations between the three entities described are discussed and the appearance of pituitary hypofunction in patients with the empty sella syndrome is emphasized.

A variant chromosome no. 21 consisting of two stalks and two satellites in tandem was detected during a survey of a human isolate. The variant segregated in three generations of a large kindred. One male had the variant no. 21, a metacentric Y, and a 47,XXY complement; however, no other evidence of chromosomal nondisjunction was found. Computer-aided analysis of sequentially stained variant no. 21 chromosomes indicated that silver-stained material corresponded to the proximal stalk region (as defined by Giemsa), but often covered both the distal stalk and satellite (also as defined by Giemsa). These data support the hypothesis that human nucleolar organizers are localized to the stalks of acrocentric chromosomes.

We studied the effect of partial retro-inverso modification of selected peptide bonds of N-terminal tetrapeptide analogues of dermorphin (H-Tyr-D-Ala-Phe-Gly-Tyr-Pro-Ser-NH2). Among the 14 compounds synthesized and tested for opioid activity, some tetrapeptides have the C-terminus carrying different amide moieties; retromodifications concern the Phe-Gly bond (Ia-f) and/or the C-terminal carboxamide function (IIIa-d, IIa-d). All pseudotetrapeptide derivatives showed opioid activity in vitro and in vivo. The most potent compounds (II) have a biological potency comparable with that of the original tetrapeptides in the guinea pig ileum preparation and in the mouse tail-flick test after intracerebral or subcutaneous administration.

Cancer mortality (1965--77) among 12,652 members of an inbred human religious isolate, the Hutterites, was compared with expectations based on mortality rates for the U.S. white population in 1970. Overall, Hutterites had significantly fewer deaths from cancer than expected (P < 0.01), due primarily to fewer lung cancers among males. Smoking is prohibited for this religious group. The most frequent types of cancers were leukemia and cancers of the digestive system, the prostate gland, and the female breast. Preliminary results suggest an association between recessive alleles and childhood leukemia. More stomach and rectal cancers were observed than expected, but differences were generally not significant. Familial aggregates of cancers of the stomach and breast are being investigated. The low frequency of cervical cancer is consistent with current evidence for an association of cervical cancer with early age at first intercourse and promiscuity, neither of which is characteristic of this population.

The structure and origin of giant nuclei in human cancer cells were studied in two cases of squamous cell carcinoma of the cervix. Feulgen-squash preparations of untreated biopsies showed the following types of nuclei: fairly evenly stained nuclei, nuclei with distinct chromocenters from which individual chromosomes sometimes seemed to radiate, and nuclei with banded structures which we tentatively interpret as some type of "polytene" chromosomes possibly showing prophase chromomeres. In neither tumor did either the small or large nuclei display any X chromatin bodies. The main processes giving rise to the different types of nuclei are endoreduplication, true endomitosis, and a kind of polytenization. The multichromosome chromocenters that fell apart possibly represent polytene chromosomes separating into individual chromosomes.

A patients with seizures, Von Willebrand disease, and symptoms of Turner syndrome was a chromosomal mosaic. In blood culture (1974), 56% of the cells were 45, X 33% 46, XXp+ and 11% 47,XXp + Xp +; in the skin, no cells with 47 chromosomes were found. Presumably the Xp + chromosome arose through a break in the Q-banded dark region next to the centromere on Xp to which an Xq had been attached. The abnormal X was late-labeling and formed a larger than normal Barr body. Of the chromatin-positive fibroblasts, 18.2% showed bipartite Barr bodies, which agrees with the hypothesis that the X inactivation center lies on the proximal part of the Xq. On the basis of the structure and behavior of the bipartite bodies in the present patient, as compared to those formed by other chromosomes with two presumed inactivation centers, we propose that the dark region next to the centromere of Xp remains active in the inactive X. In cells with 45,X and 46,XY, this region has the same relative size, whereas it is significantly shorter in the active X of three females, including the present patient, with one abnormal X. We propose that this region on the active X reveals different states of activity, as reflected in its length, depending on how many other X chromosomes are in the cell.

In three women with a 46,XXq- chromosome constitution, the length of the deletion was expressed as the ratio of the remaining part of Xq to Xp c' over a + b. In one of them (KH) this ratio was 0.33, in another (GE) 0.59, and in the third (AP) the ratio fell between these values. The break in KH is more or less on the border of the Q-dark proximal region. A comparison with relevant X-autosomal translocations indicates that the X inactivation center lies near, but not at the border of, the Q-dark and the adjoining bright region (c and d).

Familial aggregates of incomplete Müllerian fusion have been reported, but the role of genetic factors has not been elucidated. In the last several years, we have fortuitously encountered three families in which several members were affected with Müllerian fusion anomalies. In two families, several members had incomplete Müllerian fusion as traditionally described. In the third family, several members had the hand-foot-genital syndrome, a rare autosomal dominant disorder characterized not only by Müllerian fusion defects but also by skeletal (hand and foot) malformations. The etiologic heterogeneity of Müllerian fusion defects is considered.

We describe the synthesis and preliminary in vitro and in vivo pharmacological tests of five endothiotetrapeptide analogues of the opioid heptapeptide dermorphin H-Tyr-D-Ala-Phe-Gly-Tyr-Pro-Ser.-NH2. The modification obtained by substituting Phet for Phe3 or Glyt for Gly4 provided further analogues with significant opioid activity.

We studied the effect of partial retro-inverso modification of selected peptide bonds of dermorphin (H-Tyr-D-Ala-Phe-Gly-Tyr-Pro-Ser-NH2. The modifications concern two consecutive peptide bonds (Phe3-Cly4-Tyr5, I) or a single one (Gly4-Tyr5-, II or Phe3-Gly4, III). All pseudoheptapeptides showed low opioid activity in the in vitro and in vivo tests. Compound III has a biological potency comparable to that of morphine but only 2-5% of original dermorphin when tested in guinea pig ileum preparation and in mice tail-flick assay after intracerebro or subcutaneous administration.

The chromosome replication pattern of a man with 49,XXXXY was analyzed using 3H-thymidine and autoradiography as well as BrdU and acridine orange. The former technique showed a highly irregular replication pattern; the latter revealed one early replicating X chromosome, and the other three more or less asynchronously replicating. Two hypotheses seem to explain best the abnormal phenotype of males with an XXXXY sex chromosome constitution: The number of the always active regions (tip of Xp) and of the possibly always active regions (the Q-dark regions on both sides of the centromere) is increased from one to four. The replication pattern of the late-replicating X chromosomes is highly asynchronous, which might affect the phenotype. The possibility that more than one X chromosome might remain active in some cells, an even more abnormal and obviously deleterious situation, is still open.