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H Green

Publications and source records attributed to H Green.

At least 91 records · Page 5Linked to original sources

The involucrin gene of the galago. Existence of a correction process acting on its segment of repeats.

The involucrin gene of the galago, a prosimian, has been cloned and sequenced. The coding region contains a segment of repeats homologous to the segment of repeats in the gene of another prosimian, the lemur, and different from the segments of repeats in the genes of higher primates. The repeats lengths in the two prosimians are similar; and except for a single duplication of a block of repeats in the lemur alone, the number of repeats is the same. However, the nucleotide consensus sequences of the repeats differ between the two species at 3 out of 39 nucleotide positions. The repeats therefore appear to have been modified by a correction process that led toward homogeneity in the repeats of each species while permitting divergence between the two species. The correction process, an example of concerted evolution, has taken place preferentially between adjacent repeats. The numerous differences between the segments of repeats of higher primates and the segments of repeats of lower animals reveal a discontinuity in the evolutionary processes acting on the gene.

Animals↗

The involucrin gene of the gibbon: the middle region shared by the hominoids.

The evolution of the anthropoid involucrin gene has resulted largely from a process of vectorial addition of short tandem repeats. The coding region of the involucrin gene of the gibbon (Hylobates lar), including the segment of repeats, has been cloned and sequenced, and its repeat structure can now be compared with that of the other hominoids. In the gibbon, as in the others, repeat additions in the past can be assigned to early, middle, and late regions of the present-day segment of repeats. All 10 repeats of the gibbon early region were completed in a common anthropoid ancestor. All 17 repeats of the gibbon middle region were completed in a common hominoid ancestor. After divergence of the gibbon lineage, eight repeats were added to the middle region of the great ape-human lineages. Seven of these are shared by two to four species, according to the order of their divergences from each other. After its divergence, the gibbon lineage added a short species-specific late region. The gibbon also possesses an incomplete repeat just 3' of the early region, the only addition in this region in any hominoid. Comparison of the number of repeats added with the number of nucleotides substituted shows an inconstant relation between the two.

Animals↗

The involucrin genes of pig and dog: comparison of their segments of repeats with those of prosimians and higher primates.

The involucrin genes of the dog and the pig have been cloned and sequenced. Like the corresponding genes of the prosimians, each contains a homologous segment of short tandem repeats at the same position in the coding region. However, the codon sequence of the repeats in the prosimians differs significantly from that of the nonprimate mammals. This evolution has been brought about by a combination of genetic modifications (selective deletions, mutations, and gene conversions). In the anthropoids, this segment of repeats was replaced by a modern one differing in location, sequence, and repeat length. In several of its properties the modern segment has continued the prosimian trend away from the nonprimates. The overall direction of the evolution of this segment has therefore been maintained even though there have been sudden changes in the evolutionary processes acting on the gene.

Animals↗

Human growth hormone in the blood of athymic mice grafted with cultures of hormone-secreting human keratinocytes.

An established line of human epidermal keratinocytes was permanently transformed by transfection with plasmid vectors bearing the human growth hormone gene under the control of two different promoters. Suitable selection permitted the isolation of clones secreting abundant growth hormone in culture. When such cultures were grafted to athymic mice, human growth hormone could be detected in the blood of the mice at concentrations in the physiological range for more than 4 weeks. Retrograde transfer from epithelial cells should be studied further as a possible means of introducing exogenous gene products into the circulation of humans.

Animals↗

Treatment of skin ulcers with cultured epidermal allografts.

Thirty-six skin ulcers in 23 patients were treated with cultured allogeneic epidermal sheets derived from neonatal foreskin. In 73% of ulcers, there was complete healing within 8 weeks, with a mean healing time of 3.3 weeks. In the other 27%, there was reduction in ulcer size of 35% to 93% by 8 weeks after grafting. In 30 painful ulcers, pain was markedly relieved within 24 hours of grafting. The healing pattern suggested that the cultured epidermal sheets acted by stimulation of host keratinocytes to divide and migrate rather than by permanent acceptance of the allograft. Of the 26 ulcers that healed within 8 weeks, 23 (88.5%) remained healed for follow-up periods of 10 to 18 months (mean 13.7 months), with an overall mean duration of healing of 13 months.

Adult↗

Restoration of growth potential in paraclones of human keratinocytes by a viral oncogene.

Human diploid keratinocytes may be divided into three clonal types with differing capacities for proliferation. The paraclone, which has the shortest life span, is limited to 15 divisions, after which all the cells undergo programmed terminal differentiation. By means of a retroviral vector, paraclones which have not completed their life span and which consist of not more than a few hundred cells can be transduced at a high frequency with DNA complementary to the 12S transcript of the adenovirus early region 1A gene. Transformation can be detected within a single cultivation by the formation of progressively growing colonies. The transformants appear to have an unlimited growth potential, and they form a disorganized epidermis when they are grafted as an epithelium onto athymic mice. These experiments clearly show that, in order to be transformed by a viral oncogene, the target cell need not be a stem cell.

Aged↗

Vectorial expansion of the involucrin gene and the relatedness of the hominoids.

In higher primates, the coding region of the gene for involucrin, an epidermal protein, is mostly composed of a recently generated (modern) segment of repeats of a sequence of 10 codons. While the rest of the coding region has evolved only by nucleotide substitutions, the modern segment has evolved by successive addition of repeats. This process has not taken place randomly; instead, the expansion of the modern segment has been progressive from 3' to 5' end, thus adding vectorially regions that have been defined as early, middle, and late. The relatedness of the human, chimpanzee, and gorilla may be analyzed with greatest sensitivity by comparing their middle regions. The chimpanzee involucrin gene is more closely related to that of the gorilla than to that of the human.

Alleles↗

Divergent evolution of part of the involucrin gene in the hominoids: unique intragenic duplications in the gorilla and human.

The gene for involucrin, an epidermal protein, has been remodeled in the higher primates. Most of the coding region of the human gene consists of a modern segment of repeats derived from a 10-codon sequence present in the ancestral segment of the gene. The modern segment can be divided into early, middle, and late regions. We report here the nucleotide sequence of three alleles of the gorilla involucrin gene. Each possesses a modern segment homologous to that of the human and consisting of 10-codon repeats. The early and middle regions are similar to the corresponding regions of the human allele and are nearly identical among the different gorilla alleles. The late region consists of recent duplications whose pattern is unique in each of the gorilla alleles and in the human allele. The early region is located in what is now the 3' third of the modern segment, and the late, polymorphic region is located in what is now the 5' third. Therefore, as the modern segment expanded during evolution, its 3' end became stabilized, and continuing duplications became confined to its 5' end. The expansion of the involucrin coding region, which began long before the separation of the gorilla and human, has continued in both species after their separation.

Animals↗

The involucrin gene of the owl monkey: origin of the early region.

A large part of the coding region of the hominoid involucrin gene is of recent origin. This part of the gene, which we have called the modern segment, contains numerous repeats of a sequence of 10 codons, created by multiple duplications some of which consist of 3-12 repeats. We have sequenced two alleles of the involucrin gene in the owl monkey and found that the involucrin gene of this species also possesses a modern segment. By comparing the modern segment of the owl monkey with that of the hominoids, we find that only a part of this segment is shared by the two species. We call this part the early region because it must have originated in a common ancestor of the anthropoids. The rest of the hominoid modern segment does not correspond to any groups of repeats in the owl monkey and was therefore created after divergence of the two lineages. As in the hominoids, the latest additions to the modern segment of the owl monkey have been in its 5' half, which possesses different duplication patterns in the two alleles. Lineage divergences within the anthropoids can be detected at different sites within the modern segment.

Animals↗

The involucrin gene of the orangutan: generation of the late region as an evolutionary trend in the hominoids.

In the evolutionary line leading to the higher primates, the coding region of the involucrin gene evolved a segment consisting of numerous repeats of a 10-codon sequence. Additions to this segment of repeats have been made successively, thus generating regions that can be defined as early, middle, and late. The involucrin gene of the orangutan (Pongo pygmaeus abelii) possesses a segment of repeats whose early region has the same repeat structure as that in other anthropoids. The middle region is not similar in repeat structure to that of all anthropoids but is similar to that of other hominoids. The late region is unique to the species; it does not correspond at all in its repeat structure to that of the human or gorilla and is much larger. The late region of the orangutan was generated by duplications of blocks of older repeats clearly belonging to the middle region. Continued duplications extending the late region are an evolutionary trend in the hominoids. The process of addition of repeats at a particular location is a more significant aspect of the evolution of involucrin than are random nucleotide substitutions; in addition, it has proceeded more rapidly.

Animals↗

Cultured epithelial autografts for giant congenital nevi.

Eight pediatric patients with giant congenital nevi confluent over 21 to 51 percent body surface area were treated by excision and grafting. The nevus was excised to the muscle fascia, and the open wound was grafted with cultured epithelial autografts and split-thickness skin grafts. The patients have been followed from 17 to 56 months. Seventeen operations were performed in the eight patients, excising a mean of 6.9 percent body surface area at each procedure. The mean duration of anesthesia was 3.7 hours, and the mean operative blood loss was 12.3 percent estimated blood volume. The mean "take" for the cultured epithelial autografts was 68 percent, and for the split-thickness skin grafts, 84 percent. Epithelialization of open wound areas adjacent to the grafts was somewhat slower for the cultured epithelial autografts than for the split-thickness skin grafts, but it led to a healed wound in all patients except one. Ten of the 17 areas grafted with cultured epithelial autografts resulted in small open wounds that required regrafting. Wound contraction under the cultured epithelial autografts and under split-thickness skin grafts was similar and depended more on the anatomic site grafted than on the type of graft employed. in 16 of 17 operations, the cultured epithelium remained as a permanent, durable skin coverage. The use of cultured epithelial autografts allowed a larger area of excision than would have been possible with split-thickness skin grafts alone and, therefore, a more rapid removal of nevus. Cultured epithelial autograft are an important new technique in the care of patients with giant congenital nevi.

Adolescent↗

Involucrin in the epidermal cells of subprimates.

The protein involucrin is a precursor of the cross-linked envelope that forms during terminal differentiation of the keratinocyte. Most of the human involucrin molecule consists of a segment of homologous repeats of a sequence of 10 amino acids. A similar segment is present in the involucrin of other higher primates, but not in lower animals. We show here that the older part of the involucrin molecule (the ancestral segment) is present in the epidermal cells of subprimates. This has been demonstrated with antisera prepared against different peptides of the ancestral segment of the human protein. No single antiserum detects involucrin of all subprimate species, but probably all involucrins can be detected using antiserum against some sequence in the ancestral segment. Although the involucrin gene has been extensively remodeled in higher primates, its origins extend lower in the animal kingdom.

Amino Acids↗

Capillary and size interrelationships in developing rat diaphragm, EDL, and soleus muscle fiber types.

The effects of maturation on the interrelationship between skeletal muscle fiber area and capillarization was investigated in specific fiber types (I, IIa, IIb, IIc) of male Wistar rats at seven developmental periods ranging from 8 to 85 days postnatal. Fiber type specific developmental properties were compared in three different muscles, the diaphragm (DIA), extensor digitorum longus (EDL), and soleus (SOL), which are known to differ widely in function. All fiber types in each of the three muscles examined exhibited large increases in area (FA), the magnitude and time course of the increase being related to both the type of fiber and the muscle in which the fiber was located. For type I fibers, areas increased from 3- to 18-fold (SOL greater than EDL greater than DIA), whereas in type IIa fibers, area increased ranged between 5- to 11-fold (SOL greater than EDL greater than DIA). Growth rates in IIb fibers were more homogeneous between muscles ranging from 11- to 14-fold. Capillarization, as indicated by the capillary contacts per fiber (CC), increased in all fiber types regardless of muscle origin. These increases ranged between 1.7- and 2.2-fold for type I fibers, between 2.4- and 2.5-fold for type IIa fibers, and between 2.0- and 3.0-fold for type IIb fibers. In general, capillary density expressed as the ratio of the number of capillary contacts divided by the fiber area (CC/FA) progressively declined in all fiber types with age. The rate of the decline in CC/FA was mediated in large part by the changes in fiber area.(ABSTRACT TRUNCATED AT 250 WORDS)

Aging↗

Necrotizing Pneumocystis carinii vasculitis associated with lung necrosis and cavitation in a patient with acquired immunodeficiency syndrome.

Pulmonary cavitation is an infrequent manifestation of Pneumocystis carinii pneumonia in patients with the acquired immunodeficiency syndrome. The pathogenesis of lung cavities associated with P carinii pneumonia has not been fully elucidated by detailed morphologic studies. This article describes a patient with the acquired immunodeficiency syndrome and P carinii pneumonia who developed roentgenographically identified lung cavitation while being treated with azidothymidine. At autopsy, necrotizing vasculitis with vascular infiltration by Pneumocystis organisms was associated with lung necrosis and cavitation. No other causes of pulmonary cavitation were identified. Thus, in patients with acquired immunodeficiency syndrome, P carinii, per se, may result in lung necrosis and cavitation, possibly through the mechanism of necrotizing angiitis.

Acquired Immunodeficiency Syndrome↗

The glutamine residues reactive in transglutaminase-catalyzed cross-linking of involucrin.

The protein involucrin, synthesized by human keratinocytes, contains 585 amino acids, largely in the form of 10 amino acid repeats, each containing glutamines in 3 conserved positions. Involucrin is a substrate for the keratinocyte transglutaminase and is labeled by the cosubstrate amine, glycine ethyl ester. Study of tryptic peptides of involucrin shows that a single glutamine (residue 496), located 89 residues from the C-terminal end, is preferentially labeled by the enzyme. Additional glutamine residues become reactive when the molecule is fragmented. The C-terminal end, isolated as a cyanogen bromide fragment of 275 residues, is labeled equally at 2 glutamine residues. The polypeptide containing residues 148 to 280 accepts practically no amine while in intact involucrin but as a free fragment is labeled at multiple glutamine residues. It is concluded that the C-terminal and N-terminal ends of the protein are directive influences in that they suppress the reactivity of a number of glutamine residues in the intact molecule, leaving one glutamine highly preferred by the transglutaminase.

Amino Acid Sequence↗

Remodeling of the involucrin gene during primate evolution.

The protein involucrin is a product of terminal differentiation in the epidermal cell and related cell types. By comparing the nucleotide sequence of the involucrin gene of the lemur with that of the human, it is clear that the gene has undergone unusual evolution in the primates. The coding region of the gene contains an ancestral segment, most of which is common to the lemur and the human, and a species-specific segment of repeats derived from the ancestral segment. Instead of the modern segment of repeats found in the human gene, the lemur gene possesses repeats derived from another sequence at a different location in the ancestral segment. The two kinds of segments of repeats probably represent alternative ways of creating a repeat structure in the involucrin molecule. The modern segment of repeats must have been created after divergence of the higher primates from the prosimians.

Amino Acid Sequence↗