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Biomedical subjects

J D Hopkins

Publications and source records attributed to J D Hopkins.

At least 19 recordsLinked to original sources

Adjunctive treatment of congenital pigmented nevi with phenol chemical peel.

The purpose of this study was a retrospective evaluation of the treatment of congenital pigmented nevi using the phenol chemical peel technique. Patients were treated with standard Baker formula in the operating room under general anesthesia or intravenous sedation with continuous electrocardiogram monitoring. A total of 20 patients were reviewed (13 girls and 7 boys, mean age 3.8 years). Eight patients had nevi located on the face, five patients had trunk lesions, and three patients had lesions on the thighs. Two patients had nevi located on both the face and the trunk, and two patients had involvement of the face, trunk, and thigh. Three of the above patients had the classic "bathing trunk" distribution of the nevi. A test area was peeled in four patients, and in five patients preoperative biopsies were performed to rule out malignancy before initiation of therapy. An average of 2.6 treatments were performed per patient. Two patients had adjunctive dermabrasion to increase the depth of peel and to contour surface irregularities. The length of follow-up ranged from 6 to 84 months with a mean of 28 months. Healing of the wounds occurred within 2 to 3 weeks postoperatively. Seventy-five percent of patients had satisfactory cosmetic improvement in the appearance of the lesions following treatment. Four patients had recurrence of the pigmentation after an initial lightening response, three of whom had their nevi subsequently excised. There was no incidence of hypertrophic scarring or cardiac and/or renal complications. There was one death from complications of leptomeningeal melanocytosis. Chemical peeling of congenital pigmented nevi is an acceptable alternative method of therapy for those lesions that are too large for excision and primary closure or for lesions in which excision would result in unacceptable scars in areas such as the face.

Adolescent↗

Hemophilic pseudotumor of the soft tissue of the hand: a case report.

A 25-year-old, right hand dominant man with a history of Hemophilia A, presented with an enlarging 2 x 3.5 cm mass on the dorsum of his dominant hand following minor trauma. Factor VIII activity level was 12 percent of normal. A diagnosis of hemophilic pseudotumor was made and the mass continued to enlarge despite medical management. Surgical intervention was carried out in order to prevent impending skin necrosis or rupture. The patient was treated by surgical excision of the lesion along with factor replacement therapy. The patient's symptoms resolved and he had no recurrence of the tumor.

Adult↗

Osteolipoma of the hand: a case report.

A 61-year-old, healthy woman presented with a six-month history of a mass on the radial volar aspect of her dominant right hand. The patient had a history of trauma to the hand six years previously. Radiographs showed a bony lesion in the index metacarpal shaft and the MRI showed a bony lesion and soft tissue mass suggestive of a parosteal lipoma. Surgical excision of the lesion revealed a lipoma overlying a bony exostosis or osteolipoma of the hand. Post-operatively, the patient's symptoms resolved and she had no recurrence of the tumor.

Bone Neoplasms↗

A follow-up study of 33 subdiagnostic eating disordered women.

Thirty-three female subjects with subdiagnostic DSM-III-R anorexia nervosa (SAN) and/or subdiagnostic bulimia nervosa (SBN) were reinterviewed 24 to 52 months (mean 41 months) after seeking treatment for an eating disorder. Subjects were administered a semistructured interview by telephone and assessed for level of functioning, eating disorder symptoms, course of illness, and treatment sought. During the course of the follow-up, 15 (46%) subjects went on to meet full DSM-III-R criteria for AN and/or BN. At follow-up, 4 (12%) met full DSM-III-R criteria for AN and/or BN, 22 (67%) were subdiagnostic, and 6 (18%) had recovered. The high percentage of subdiagnostic women that eventually develop full DSM-III-R criteria for AN and/or BN and the low rates of recovery at 2 to 4 years suggest that the current diagnostic criteria may be too restrictive.

Adolescent↗

Plasmid diversity in Escherichia coli isolated from processed poultry and poultry processors.

Plasmids of bacteria selected from different bacterial populations because they shared a distinctive antimicrobial resistance phenotype have sometimes had identical restriction fragments. Such identical plasmids are thought to belong to small and thus epidemic clones because the plasmid content of unselected resistant isolates has seemed diverse. To survey this presumed diversity and its implications for the lineage of resistance plasmids we examined the transferability, sizes and EcoR1 restriction fragment sizes of plasmids in both Escherichia coli isolated randomly from poultry raised by 16 growers as they were being processed through two plants and in isolates from the urine of women processing poultry in those plants. Forty two (24%) of 175 resistant isolates from poultry of 16 growers and 9 (26%) of 34 resistant isolates from the poultry processors transferred resistance conjugatively to varied combinations of antimicrobials. No poultry isolate had both the same expressed and the same transferred combination as any processor's isolate. The DNA bands which could be discerned in electrophoresis gels of restricted or unrestricted plasmid extracts of isolates or their transconjugants from 156 of the poultry and 24 of the poultry processors appeared diverse. Pairs of related-appearing plasmids were seen in consecutive isolates of poultry from each of two growers and in one pair from different growers. One set of identical-appearing plasmids was seen in 3 consecutive isolates from poultry of one grower, others in 2 consecutive isolates from a second grower's poultry, in 2 non-consecutive isolates of a third grower's, and in single isolates from poultry of 2 different growers. None of the plasmids from any of the human isolates appeared related to those from any other human isolate or to those of any poultry isolate. These results indicate that resistance plasmids are highly diverse and that all but two of the exceptions to complete diversity in the isolates surveyed here could be ascribed to cross colonization within flocks of individual poultry growers. Also, while none of the plasmids in the poultry isolates appeared ancestral to any of plasmids in the poultry processors' isolates, their diversity indicates that those sampled plasmids would be only a very small fraction of the total number of different plasmids in bacteria colonizing poultry processed at that time or earlier.

Animals↗

Gly-238-Ser substitution changes the substrate specificity of the SHV class A beta-lactamases.

The SHV-type beta-lactamase SHV-2A is related to SHV-1 by a Gly-238-Ser replacement. Strains carrying SHV-2A are resistant to the third generation cephems cefotaxime and ceftizoxime, whereas those that carry SHV-1 are sensitive to these drugs. We present a kinetic analysis of a SHV-1 and SHV-2A enzymes, with the goal of gaining insight into the role of residue 238 in hydrolyzing cefotaxime and ceftizoxime. SHV-2A shows altered kinetic properties for a number of other cephems that also have heterocyclic side chains at the amino position of the 7-aminocephalosporanic acid nucleus (R1 side chain), including a significantly higher kcat/Km than does SHV-1 for cephaloridine, cephalothin, and cefotiam. Two cephems with straight chain R1 substitutions, cephalosporin C and cephacetrile, are not hydrolyzed more efficiently by SHV-2A. These results indicate that the Ser-238-Gly substitution increases the affinity toward cephems with a heterocyclic ring in the R1 side chain. In addition, the data for ampicillin and benzylpenicillin show that addition of a nitrogen to the second carbon of the R1 side chain of a penem results in a lower kcat/Km for SHV-2A relative to SHV-1. These data strongly suggest that the previously proposed hydrogen bond formation between Ser-238 and the second carbon nitrogen of cefotaxime is not an important factor in hydrolysis by SHV-2A. We propose that the Gly-238 to Ser-238 replacement in SHV-2A has altered the hydrophobic pocket so that it can better accommodate cephems with bulky R1 side chains.

Amino Acid Sequence↗

Evolved neomycin phosphotransferase from an isolate of Klebsiella pneumoniae.

A new aminoglycoside resistance gene (aphA1-IAB) confers high-level resistance to neomycin. The sequence of aphA1-IAB is closely related to aphA1 found in the transposons Tn4352, Tn903 and Tn602. For example, aphA1-IAB differs from aphA1-903 at five nucleotides that result in four amino acid replacements. The enzyme encoded by aphA1-IAB has a significantly higher turnover number with neomycin, kanamycin and G418 as substrates than does the aphA1-903 enzyme. A parsimonious phylogenetic tree suggests that aphA1-IAB evolved from an ancestral form that is closely related or identical to the aphA1 found in Tn903. The excess of replacement substitutions over silent substitutions in aphA1-IAB, as well as its convergence toward aphA3 from Staphylococcus aureus, is indicative of selective evolution. Our hypothesis to explain these results is that aphA1-IAB evolved under the selective pressure of neomycin use in relatively recent times.

Amino Acid Sequence↗

Nosocomial spread of an amikacin resistance gene on both a mobilized, nonconjugative plasmid and a conjugative plasmid.

Resistance to amikacin among members of the family Enterobacteriaceae at a hospital in Venezuela rose from 2% in 1979 to 5% in 1984 and 10% in 1985 as amikacin usage rose 20-fold to exceed gentamicin usage. Resistance to gentamicin remained at 25 to 27%. We examined the plasmids from 21 isolates obtained in 1984 and 1985. Nine of eleven in 1984 and three of ten in 1985 carried aacA and sul on a 3.8-kb BamHI fragment of pBWH300, a 10.4-kb nonconjugative plasmid that had been mobilized into strains of six species by at least two different coresident conjugative plasmids. Six 1985 isolates of two species carried these genes on a similar BamHI fragment of the 104-kb conjugative plasmid pBWH303. One isolate in 1984 and one in 1985 carried the 69-kb conjugative plasmid pBWH301, which had aacA as the promoter-proximal gene of an operon that also encompassed the cat and aadB resistance genes. Another conjugative plasmid, pBWH302, was found in a single isolate. It carried a different aacA allele on the functional transposon Tn654, which appeared to be closely related to Tn1331, a transposon previously isolated in Argentina and Chile. Increased selection may thus have led to dissemination of an endemic aacA allele on two endemic plasmids, one spread by mobilization, with occasional intrusion of additional aacA alleles from outside.

Alleles↗

Direct involvement of IS26 in an antibiotic resistance operon.

The plasmid pBWH77, originally found in an isolate of Klebsiella pneumoniae, harbors a new antibiotic resistance operon containing two resistance genes transcribed from an IS26-hybrid promoter, as shown by nucleotide sequencing, mRNA mapping, and the effect of inserting a transcription terminator within the promoter-proximal gene. The nucleotide sequence of this region revealed that the operon (IAB) is made up of three sections that are closely related to previously described genetic elements. The -35 region of the promoter, together with the adjacent sequence, is identical to sequences of the IS26 element. One of the resistance genes, aphA7, which is located next to the hybrid promoter, confers assistance to neomycin and structurally related aminoglycosides. This aphA7 gene is highly homologous to aphA1 of Tn903, with five nucleotide differences. The second gene, blaS2A, encodes an evolved SHV-type beta-lactamase with a pI of 7.6 that confers resistance to the broad-spectrum cephalosporins cefotaxime and ceftizoxime. The deduced amino acid sequence of SHV-2A shows that amino acid 238 is a serine, a residue reported to confer resistance to cefotaxime. We discuss how the operon may have evolved by a combination of insertion sequence-mediated genetic rearrangements and acquisitive evolution. Using phylogenetic parsimony, we show that aphA7 in the IAB operon evolved from an ancestral form similar to aphA1 in Tn903 and that blaS2A evolved from an ancestral form similar to blaS1.

Base Sequence↗

Bilateral ectopic pheochromocytomas: computed tomographic and sonographic correlation.

We describe a case of clinically unsuspected bilateral ectopic pheochromocytomas in a young woman. The bilateral retroperitoneal masses were located in the lower sympathetic ganglia at the level of the organ of Zuckerlandl. Computed tomography (CT) accurately identified the location and vascular characteristics of these neoplasms and provided access for percutaneous biopsy. Sonographic correlation is also presented.

Adult↗

Functional and structural map of pLST1000: a multiresistance plasmid widely distributed in Enterobacteriaceae.

pLST1000, an 80-kb plasmid found in Enterobacteriaceae in North and South America, harbors the aadB and several other resistance genes. We suggested earlier that, because of its widespread distribution, pLST1000 could act as a carrier plasmid, bringing the aadB gene to new locations. This paper presents the restriction enzyme recognition site and functional map of the plasmid. The resistance genes lie in a discreet region. The aadB and aadA genes form an operon with the aadB gene promoter proximal. This operon is flanked by bla-TEM and bla-OXA2 genes, the former located in a functional Tn3-like transposon. This arrangement is similar to that of relatives of the transposon TN21, where additional resistance genes are precisely inserted in recombinational "hot spot" sequences that flank the aadA gene. We were not able to demonstrate transposition of the aadB gene in Escherichia coli. A sul gene and mer operon lie beyond the bla-OXA2 gene. The transfer genes form a single region, defined by insertions of Tn5-132 that give the Tra- phenotype.

Drug Resistance, Microbial↗

Stimulation of precise excision and recombination by conjugal proficient F'plasmids.

Large F plasmids such as F'128 stimulate precise excision of the transposons Tn5 and Tn10 in E. coli K12. This stimulation occurs when the transposons are either on the F'128 plasmid or the bacterial chromosome. Stimulation of precise excision is dependent upon conjugal transfer proficient F'plasmids. Tra- mutations which are defective in conjugal transfer negate this F'128 plasmid stimulation effect. F'128 traS mutations, which are surface exclusion defective and thus permit matings between male cells, thereby increasing conjugal transfer, increase the F plasmid stimulation effect. When the F' plasmid is present in a cell with the small plasmid, pRS31, carrying the traS to traZ region of F, stimulation of precise excision is no longer observed. This complementation-like activity by pRS31 is abolished by a Tn5 insertion in the traS gene. Data are presented supporting the notion that F' plasmid stimulation of precise excision occurs in the recipient during conjugal transfer. F'128 traS also stimulates recA-dependent recombination between DNA sequences on the small, nontransferrable plasmid pRDK41, DNA sequences that are unrelated to those of the F plasmid. The F'plasmid stimulation of precise excision of Tn5 is not seen with F+ but only with certain F's with large insertions of chromosomal DNA.

Conjugation, Genetic↗

Molecular evolution, species distribution, and clinical consequences of an endemic aminoglycoside resistance plasmid.

During the first 6 years after appearing in one hospital, a 92-kilobase conjugative plasmid, pBWH1, which encoded resistance to chloramphenicol and sulfonamides and determined TEM-1 beta-lactamase and 2''-aminoglycoside nucleotidyltransferase, underwent a variety of molecular changes. It was most prevalent initially in isolates of Klebsiella pneumoniae, then in isolates of Serratia marcescens, and finally, after nearly disappearing, in isolates of Enterobacter cloacae. Evolutionary changes in the plasmid did not account for its shifts in species distribution, since the original molecule was found in isolates of each species. The late resurgence of pBWH1 occurred after a copy of its original molecule entered a distinctive ornithine decarboxylase-negative strain of E. cloacae, new to the hospital. The resulting transconjugant strain, chromosomally resistant to topical silver salts and to cephalosporins, and with the addition of pBWH1-encoded aminoglycoside resistance, spread in the hospital by causing an outbreak of sepsis in the burn unit, where these were commonly used antibacterial agents. Thus, an endemic plasmid became prevalent in a new host species because one of its genes supplemented the fitness of an uncommon strain of the species for a particular clinical niche.

Aminoglycosides↗

Genetic and physical characterization of IncM plasmid pBWH1 and its variance among natural isolates.

We present a genetic and physical characterization of the IncM plasmid pBWH1. A physical map was constructed for the enzymes EcoRI, BamHI, SalI, BglII, HindIII, MstII, and XhoI. A series of deletions and a series of subclones of pBWH1 were constructed and used to determine the locations on this map of the transfer region; the replication region; and the genes determining resistance to beta-lactams, chloramphenicol, the sulfonamides, and gentamicin. We compared 51 different isolates, including isolates which had lost individual antibiotic resistances or the transfer phenotype, and showed that variations occurred in all regions of the plasmid genome. Frequently, correlations could be made between phenotypic variation and variation of the EcoRI fragments which contained the gene determining that phenotype.

Chromosome Mapping↗

Intercontinental spread of a new antibiotic resistance gene on an epidemic plasmid.

Bacteria of different genera isolated at nine medical centers in different parts of the United States and at one center in Venezuela during the first decade of gentamicin usage carried the gentamicin resistance gene 2"-aminoglycoside nucleotidyltransferase on the same transferable plasmid. Such widespread dissemination of a newly observed resistance gene on one plasmid suggests that a new resistance gene may emerge once on a single plasmid, which then carries it to other centers and other plasmids. The resistance gene might, therefore, be contained if detected early.

Bacteria↗

Resistance to antibiotics in clinical isolates of Klebsiella pneumoniae.

The antibiotic resistance of Klebsiella pneumoniae isolates from 12 medical centers worldwide, over a 1- to 6-year period, were tested. Clinical isolates of K. pneumoniae were resistant to ampicillin and carbenicillin. Resistance to other antibiotics was less frequent with isolates of K. pneumoniae from 5 of 6 US centers than with those from 6 centers outside the US. In nearly all of the centers, resistance to sulfamethoxazole-trimethoprim, gentamicin, tobramycin, or chloramphenicol was more frequent in isolates of K. pneumoniae than in those of Escherichia coli, while the reverse was true for resistance to tetracycline. Resistance to multiple antibiotics declined gradually in isolates of K. pneumoniae at one center, but rose abruptly again with dissemination of a new plasmid.

Ampicillin↗

Trimethoprim resistance in multiple genera of Enterobacteriaceae at a U.S. hospital: spread of the type II dihydrofolate reductase gene by a single plasmid.

The percentage of clinical isolates of several species of Enterobacteriaceae, particularly Escherichia coli and Klebsiella pneumoniae, resistant to trimethoprim (TMPR) has increased gradually at the Brigham and Women's Hospital (Boston) in recent years. Thirty-seven of 42 TMPR isolates from six species of gram-negative bacilli conjugally transferred TMP resistance to K12 E. coli. beta-Lactam resistance cotransferred from 21 of the 37 donors, and sulfamethoxazole (SMZ) resistance cotransferred from five of the 37 donors. Plasmids that encoded TMP resistance either alone or with SMZ resistance had a molecular size of approximately 52.5 kilobases, with identical restriction endonuclease-generated "fingerprints." Plasmids encoding beta-lactam-mediated resistance (beta R) were approximately four kilobases larger and had fragment patterns that were identical for all of the TMPR/beta R plasmids tested and had many restriction endonuclease-generated bands in common with TMPR plasmids. Radiolabeled dihydrofolate reductase (DHFR) probes identified the type II DHFR as the determinant of TMP resistance. In contrast with reports from Europe, TMP resistance in multiple species of Enterobacteriaceae was found to be spread in one hospital by a single, stable conjugative plasmid that has a wide host range and encodes the type II DHFR gene.

Conjugation, Genetic↗

Diagnostic microbiology laboratory susceptibility test results discriminate distinctive antibiotic resistance plasmids.

The sizes of the zones of inhibition around routinely tested antibiotic disks classified gentamicin-resistant isolates of Klebsiella pneumoniae from one hospital into four major antibiotype classes. From each isolate of the prevalent class (A1), two plasmids could be transferred conjugally. One carried genes for resistance to tetracycline, sulfonamides, and chloramphenicol, and for the SHV beta lactamase. The other carried genes for two aminoglycoside-inactivating enzymes, APH (3')-I and AAC (3)-III, for the TEM 1 beta lactamase, and for resistance to sulfonamides. Transconjugants of either plasmid from any A1 isolate yielded the same DNA fragments after restriction endonuclease digestion, but the two plasmids had no fragments in common. Fragments or genes from either plasmid were variously combined or lacking in plasmids from variant isolates (A2, A3, and A4). Plasmids transferable from isolates of classes B and C shared no common DNA restriction fragments with each other or with either plasmid from Class A. Fragments and genes of the plasmids from C isolates, however, were identical with those of a plasmid endemic in a nearby hospital. Routine monitoring by diagnostic microbiology laboratories of distinctive antibiotypes and of the plasmids that produce them would aid infection control and antibiotic usage policy.

Anti-Bacterial Agents↗