Child-health nurses' experiences from using pictorial support with families within child-health services in Sweden.

AIM: To gain insight into child-health nurses' experiences of using pictorial support in health visits within child-health services. METHODS: A qualitative study involving interviews conducted with 17 child-health nurses in Sweden. The interview data were analysed using content analysis. RESULTS: The nurses experienced that pictorial support could facilitate communication with families and increase opportunities for children to participate in child-health services, although it may come with challenges. This theme can be broken down into three main categories: (1) Pictorial support makes interaction with families clearer and easier and is used in different ways; (2) The design and extensiveness of the pictorial support can create obstacles; and (3) Pictorial support influences children's attitudes towards, and participation in, health visits. CONCLUSION: Pictorial support is an important and useful tool in child-health nurses' own work and improves their communication with children and caregivers during health visits. It can also increase children's participation and help them express themselves. Communicative tools such as pictorial support are very helpful to healthcare professionals striving to offer child- and family-centred care.

Persisting speech difficulties at 7-8 years of age - a longitudinal study of speech production in internationally adopted children with cleft lip and palate.

AIM: To longitudinally investigate speech production (consonant proficiency, consonant errors and perceived velopharyngeal competence) in 17 internationally adopted (IA) children with unilateral cleft lip and palate (UCLP) at three time points. METHOD: Consonant proficiency (percent consonants correct, consonant inventory) and number and type of consonant errors were calculated based on blind phonetic transcriptions of words from the Swedish Test of Articulation and Nasality (SVANTE). Velopharyngeal competence was perceptually rated by three blinded experienced speech-language pathologists at the ages of 3, 5 and 7-8 years. RESULTS: A significant positive development of speech production was found, although most children still scored very low for consonant proficiency at the age of 7-8 compared with normative values: the median for percent consonants correct was 79.7 and many children still had persisting cleft-related and developmental consonant errors. At the age of 7-8, almost half of the children were rated as having a competent velopharyngeal function and only three as having an incompetent velopharyngeal function. CONCLUSION: Persisting speech difficulties at school age in IA children with UCLP were found in the present study, which is one of the very few longitudinal studies. Our results highlight the need for detailed follow-up of speech production in clinical settings. Speech disorders may have a severe impact on a child's intelligibility and participation with peers, and there is a need for more studies investigating the actual everyday effect of the difficulties found.

Facing an unexpected reality - oscillating between health and suffering 4-6 years after bariatric surgery.

BACKGROUND: Obesity is a significant public health problem that is on the increase worldwide, and treatment with bariatric surgery is becoming more and more common. This type of surgery has proved to be good for weight reduction and for preventing complications, but few studies have investigated patients' long-term experiences of health and suffering. AIM: To explore people's experiences of health after bariatric surgery. What are their thoughts about their life, body and sexuality? METHODS: This study is based on semi-structured interviews with eight women and eight men, 4-6 years after bariatric surgery. The data were analysed using qualitative content analysis and resulted in 5 main themes and 14 subthemes. RESULTS: The new body enabled a healthy life due to better treatment in society, enhanced self-esteem, the pleasure of purchasing clothes and the courage to become more sexually active. At the same time, the body could be experienced as so unfamiliar that their life was dominated by despondency, a lack of freedom and a feeling of being lost, which made them wish to return to their old body. CONCLUSION AND IMPLICATIONS: The participants received extensive information before as well as follow-up conversations up to one year after surgery. Nevertheless, they all experienced that changing from life as an obese person to a radically reduced body often meant a confrontation with an unexpected reality that oscillated between health and suffering. This indicates that preparedness for the life changes that bariatric surgery may entail is inadequate and that moving towards health and suffering takes its own time. Therefore, more time should be allocated to talking about how life is and can become in the long term, which may facilitate a dialogical, person-centred approach to the setbacks and situations each person needs to manage in order to improve her/his health.

The physical and mental impact of surviving sepsis - a qualitative study of experiences and perceptions among a Swedish sample.

BACKGROUND: Sepsis is a critical illness with high morbidity and mortality rates. Each year, sepsis affects about 48.9 million people all over the world. This study aims to illuminate how sepsis survivors experience sepsis and the impact of sepsis, as well as the health-related quality of life thereafter. METHODS: An interview study with eight sepsis survivors was carried out in Sweden with an inductive qualitative method. The data were analyzed with content analysis. RESULTS: Four themes were identified during the analysis; The experience of health care and being a sepsis patient, New circumstances´ impact on life, Family and social interactions, and The psychological impact on life. The lack of information about how sepsis can impact the survivors' lives and what to expect can lead to prolonged agony. The long recovery time comes as an unexpected and unpleasant surprise to those affected. Initially, the sepsis survivors are almost euphoric that they have survived, which can later lead to chock and trauma when they realize that they could have died. This insight needs to be processed in order to reach reconciliation with life after sepsis. CONCLUSION: Sepsis has a huge impact on both physical and mental aspects of life. Many survivors suffer from persistent residual symptoms of varying degrees, to which they have to adapt. The sepsis survivors need individually adjusted information about the sepsis recovery trajectory, and what to expect during and after the hospital stay.

Internationally adopted children with and without a cleft lip and palate showed no differences in language ability at school-age.

AIM: The aim of this study was to investigate language ability in internationally adopted children aged 7-8 years with and without a unilateral cleft lip and palate. METHODS: We compared 27 internationally adopted children with a unilateral cleft lip and palate, adopted from China, with a group of 29 children without a cleft lip and palate, adopted from different countries. Participants were recruited from two cleft lip and palate teams in Sweden and through adoption organisations. Assessments were performed using standardised tests of speech and of receptive and expressive language ability. In addition, a parental questionnaire in which speech, language and communication aspects were rated was used. RESULTS: There were no significant differences in language ability between the groups. The only difference was related to speech ability, where the internationally adopted children with unilateral cleft lip and palate scored significantly lower. However, a high proportion of children in both groups scored low on measures of expressive language compared with test norms. CONCLUSION: The results suggested that having a cleft lip and palate did not increase the risk of language difficulties. Instead, being internationally adopted may be associated with a risk of delayed language development lasting for several years post-adoption.

Internationally Adopted Children With Unilateral Cleft Lip and Palate-Consonant Proficiency and Perceived Velopharyngeal Competence at the Age of 5.

OBJECTIVE: To compare consonant proficiency, consonant errors, and the perceived velopharyngeal (VP) competence in internationally adopted (IA) children with unilateral cleft lip and palate (UCLP) and nonadopted (NA) children with the same cleft-palate type at age 5. DESIGN: Case-control study based on phonetic transcriptions of standardized speech recordings of 5-year-olds at a tertiary hospital. PARTICIPANTS: Twenty-five IA children were compared to 20 NA children. All consecutive patients at a cleft lip and palate center participated. MAIN OUTCOME MEASURE(S): Consonant proficiency was measured using percentage consonants correct, percentage consonants correct-adjusted for age, percentage correct place, percentage correct manner, and consonant inventory. Cleft speech characteristics (CSCs), developmental speech characteristics (DSCs), and the perceived VP competence were also measured. RESULTS: The IA children had significantly lower values for all consonant proficiency variables (p < .05) and a smaller consonant inventory (p = .001) compared to the NA children. The IA children had a higher frequency of CSCs (IA = 84%, NA = 50%, p < .05) and DSCs (IA = 92%, NA = 65%, p = .057), and twice as many IA children as NA children had perceived VP incompetence (IA = 52%, NA = 25%, p = .17). CONCLUSIONS: Severe speech disorder was more common in IA children than in NA children at age 5. Most importantly, the speech disorders seem to be not only cleft-related. More detailed speech assessments with a broader focus are needed for IA children with UCLP. Longitudinal studies are recommended to further investigate the impact of speech difficulties in IA children's daily lives.

Speech production in 3-year-old internationally adopted children with unilateral cleft lip and palate.

BACKGROUND: In the last decade, a large number of children with cleft lip and palate have been adopted to Sweden. A majority of the children were born in China and they usually arrive in Sweden with an unoperated palate. There is currently a lack of knowledge regarding speech and articulation development in this group of children, who also have to deal with a late first language switch. AIMS: To study consonant proficiency in 3-year-old internationally adopted children with unilateral cleft lip and palate (UCLP) compared with peers with UCLP born in Sweden. Also to study the type and frequency of consonant errors and to perceptually compare velopharyngeal competence between the groups. METHODS & PROCEDURES: Thirty-two children born between 2006 and 2010 with UCLP participated in the study-14 adopted from China and 18 children born in Sweden. Both groups were treated by the same cleft palate team. Audio recordings at 3 years of age were perceptually analysed by blinded listeners. Consonant proficiency was measured via per cent consonants correct adjusted for age (PCC-A), per cent correct manners (PCM) and per cent correct places (PCP). The prevalence of audible nasal air leakage and velopharyngeal competence were judged and compared between groups. The type and frequencies of consonant errors related to place and manner of articulation were also analysed. OUTCOMES & RESULTS: The internationally adopted children had significantly fewer correct consonants compared with the Swedish-born children. This was true for PCC-A, PCP and PCM. This group also had significantly higher prevalence of glottal stops/fricatives and deleted target consonants more often. Also the internationally adopted children had a higher prevalence of incompetent velopharyngeal function. The only outcome variable with similar results in the groups was audible nasal air leakage. CONCLUSIONS & IMPLICATIONS: The present study indicated that there were significant differences regarding consonant proficiency and velopharyngeal competence between internationally adopted children with a UCLP and their Swedish-born peers with UCLP at the age of 3 years. Internationally adopted children with UCLP should be considered an at risk group for a higher prevalence of speech difficulties than non-adoptees. Thus, it is particularly important to follow this group of children over time. Longitudinal studies of speech and language development in internationally adopted children with UCLP are needed.

Similar alteration of motor unit recruitment strategies during the anticipation and experience of pain.

A motor unit consists of a motoneurone and the multiple muscle fibres that it innervates, and forms the final neural pathway that influences movement. Discharge of motor units is altered (decreased discharge rate and/or cessation of firing; and increased discharge rate and/or recruitment of new units) during matched-force contractions with pain. This is thought to be mediated by nociceptive (pain) input on motoneurones, as demonstrated in animal studies. It is also possible that motoneurone excitability is altered by pain related descending inputs, that these changes persist after noxious stimuli cease, and that direct nociceptive input is not necessary to induce pain related changes in movement. We aimed to determine whether anticipation of pain (descending pain related inputs without nociceptor discharge) alters motor unit discharge, and to observe motor unit discharge recovery after pain has ceased. Motor unit discharge was recorded with fine-wire electrodes in the quadriceps of 9 volunteers. Subjects matched isometric knee-extension force during anticipation of pain (anticipation: electrical shocks randomly applied over the infrapatellar fat-pad); pain (hypertonic saline injected into the fat-pad); and 3 intervening control conditions. Discharge rate of motor units decreased during pain (P<.001) and anticipation (P<.01) compared with control contractions. De-recruitment of 1 population of units and new recruitment of another population were observed during both anticipation and pain; some changes in motor unit recruitment persisted after pain ceased. This challenges the fundamental theory that pain-related changes in muscle activity result from direct nociceptor discharge, and provides a mechanism that may underlie long-term changes in movement/chronicity in some musculoskeletal conditions.

Prostaglandin D2 induces contractions through activation of TP receptors in peripheral lung tissue from the guinea pig.

Prostaglandin D(2) (PGD(2)), released through mast cell activation, is used as a non-invasive biomarker in patients with asthma. Since PGD(2) can elicit opposing effects on airway tone via activation of the PGD(2) receptors DP(1) and DP(2) as well as the thromboxane receptor TP, the aim of this study was to characterize the receptors that are activated by PGD(2) in the guinea pig lung parenchyma. PGD(2) and the thromboxane analog U46619 induced concentration-dependent contractions. U46619 was more potent and caused stronger effect than PGD(2). The specific TP receptor antagonist SQ-29548 and the combined TP and DP(2) receptor antagonist BAYu3405 concentration-dependently shifted the curves for both agonists to the right. The DP(1) receptor agonist BW245 induced a weak relaxation at high concentrations, whereas the DP(1) receptor antagonist BWA868C did not affect the PGD(2) induced contractions. The specific DP(2) receptor agonist 13,14-dihydro-15-keto-PGD(2) showed neither contractile nor relaxant effect in the parenchyma. Furthermore, studies in precision-cut lung slices specified that airways as well as pulmonary arteries and veins contracted to both PGD(2) and U46619. When the lung parenchyma from ovalbumin sensitized guinea pigs were exposed to ovalbumin, both thromboxane B(2) and PGD(2) were released. Ovalbumin also induced maximal contractions at similar level as PGD(2) in the parenchyma, which was partly reduced by SQ-29548. These data show that PGD(2) should be recognized as a TP receptor agonist in the peripheral lung inducing contraction on airways, arteries and veins. Therefore, a TP receptor antagonist can be useful in combination treatment of allergic responses in asthma.

Duration of methicillin-resistant Staphylococcus aureus colonization after diagnosis: a four-year experience from southern Sweden.

BACKGROUND: The duration of colonization with methicillin-resistant Staphylococcus aureus (MRSA) is not well known and there is debate as to whether a patient colonized with MRSA ever can be defined as 'MRSA-negative'. METHODS: Since 2003 all notified MRSA cases have been systematically followed in Skåne County, southern Sweden. Cultures are taken from the nares, throat, perineum and possible skin lesions. Contact tracing is conducted. The screening program continues as long as cultures are positive and then until 1 y of consecutive negative cultures for MRSA is completed. RESULTS: Of the 578 MRSA cases during 2003-2006, 535 were included in this retrospective study. The median duration of colonization with MRSA was 5.9 months. Having household contacts with MRSA, young age, spa-type t002 and colonization in 2 or more locations, were significantly associated with a longer duration of colonization. Having a clinical infection treated with antibiotics (compared to clinical infection with no antibiotic treatment or asymptomatic carriage) was significantly associated with a shorter carriage time. Eradication treatment was associated with a shorter carriage time. CONCLUSION: These results may have implications for the management of patients with MRSA carriage. The study indicates that MRSA carriage can be defined as 'negative' in a follow-up program and shows the importance of performing contact tracing among household members.

Parents' experiences of an abnormal ultrasound examination - vacillating between emotional confusion and sense of reality.

BACKGROUND: An ultrasound examination is an important confirmation of the pregnancy and is accepted without reflection to any prenatal diagnostic aspects. An abnormal finding often comes unexpectedly and is a shock for the parents. The aim was to generate a theoretical understanding of parents' experiences of the situation when their fetus is found to have an abnormality at a routine ultrasound examination. METHODS: Sixteen parents, mothers and fathers, whose fetus had been diagnosed with an abnormality during an ultrasound scan in the second or third trimester, were interviewed. The study employed a grounded theory approach. RESULTS: The core category vacillating between the emotional confusion and sense of reality is related to the main concern assessment of the diagnosis impact on the well-being of the fetus. Two other categories Entering uncertainty and Involved in an ongoing change and adaptation have each five sub-categories. CONCLUSIONS: Parents are aware of that ultrasound examination is a tool for identifying abnormalities prenatally. The information about the abnormality initially results in broken expectations and anxiety. Parents become involved in ongoing change and adaptation. They need information about the ultrasound findings and the treatment without prolonged delay and in a suitable environment. The examiner who performs the ultrasound examination must be aware of how anxiety can be intensified by environmental factors. All parents should to be offered a professional person to give them support as a part of the routine management of this situation.

Molecular evolution of Theta-class glutathione transferase for enhanced activity with the anticancer drug 1,3-bis-(2-chloroethyl)-1-nitrosourea and other alkylating agents.

Glutathione transferase (GST) displaying enhanced activity with the cytostatic drug 1,3-bis-(2-chloroethyl)-1-nitrosourea (BCNU) and structurally related alkylating agents was obtained by molecular evolution. Mutant libraries created by recursive recombination of cDNA coding for human and rodent Theta-class GSTs were heterologously expressed in Escherichia coli and screened with the surrogate substrate 4-nitrophenethyl bromide (NPB) for enhanced alkyltransferase activity. A mutant with a 70-fold increased catalytic efficiency with NPB, compared to human GST T1-1, was isolated. The efficiency in degrading BCNU had improved 170-fold, significantly more than with the model substrate NPB. The enhanced catalytic activity of the mutant GST was also 2-fold higher with BCNU than wild-type mouse GST T1-1, which is 80-fold more efficient than wild-type human GST T1-1. We propose that GSTs catalyzing inactivation of anticancer drugs may find clinical use in protecting sensitive normal tissues to toxic side-effects in treated patients, and as selectable markers in gene therapy.

Parents' worried state of mind when fetal ultrasound shows an unexpected finding: a comparative study.

OBJECTIVE: Most parents yearn for a second-trimester ultrasound examination and feel excitement about it, but some also worry about what the examination will show. According to prior research, using only generic instruments or specific questionnaires, anxiety decreases when the ultrasound findings are normal. The aim of this study was to compare parents' worry (Parents' Expectations, Experiences, and Reactions to Ultrasound [PEER-U] State of Mind Index) and sense of coherence before and after a routine second-trimester ultrasound examination when it showed normal or abnormal findings. METHODS: A 1-year cohort study was performed at a Swedish university hospital. A total of 2049 parents who had their second-trimester ultrasound examinations there filled in a questionnaire consisting of 2 parts before and after the examinations. RESULTS: Parents with normal ultrasound findings were less worried than parents with abnormal findings. The group with normal findings also showed less worry after the examination than before. A sex analysis showed similar patterns. CONCLUSIONS: Parents with abnormal ultrasound findings are more worried and anxious. The new instrument, the PEER-U State of Mind Index, not only measures parents' worry but can also expose what influences their ultrasound examination experience.

Specific mediator inhibition by the NO donors SNP and NCX 2057 in the peripheral lung: implications for allergen-induced bronchoconstriction.

BACKGROUND: The aim of this study was to examine potential therapeutic effect of the two NO donors NCX 2057 (3-(4-hydroxy-3-methoxyphenyl)-2-propenoic acid) 4-(nitrooxy)butyl ester) and SNP (sodium nitroprusside) on the early allergic airway response in the peripheral lung. METHODS: The experiments were performed in guinea pig lung parenchyma (GPLP) derived from ovalbumin (OVA) sensitized guinea pigs. The effects of NCX 2057 and SNP were evaluated by contractile responses and mediator release during OVA challenge. The generation of nitrite and nitrate was assessed by chemiluminescence. Statistical analysis was evaluated by ANOVA. RESULTS: Cumulatively increasing concentrations of OVA (1-10,000 ng/ml) induced concentration-dependent contractions of the GPLP that were reduced by NCX 2057 (100 microM, p < 0.001) and SNP (100 microM, p < 0.05). Antigen-induced eicosanoid release was decreased by NCX 2057 (100 microM, p < 0.001) but not by SNP (100 microM), whereas the release of histamine was reduced by SNP (100 microM, p < 0.001) but not by NCX 2057 (100 microM). In addition, NCX 2057 (0.1-100 microM), but not SNP (0.1-100 microM), relaxed leukotriene D4 (10 nM) precontracted GPLP (p < 0.01). The guanylyl cyclase inhibitor ODQ had no effect on the NCX 2057 mediated relaxation. SNP released significantly less nitrite than NCX 2057. CONCLUSION: Although both SNP and NCX 2057 reduced the release of pro-inflammatory mediators, their profiles were distinctly different. Furthermore, NCX 2057 also induced smooth muscle dilation in the GPLP. The findings point to specific anti-inflammatory effects of different NO donors in the peripheral lung tissue.

Parental level of anxiety, sense of coherence and state of mind when choroid plexus cysts have been identified at a routine ultrasound examination in the second trimester of pregnancy: a case control study.

The aim of the study was to compare parents' experience of a routine ultrasound examination in the second trimester, when a choroid plexus cyst/cysts (CPC) were found (Study group; n = 22), with matched controls where no fetal deviations were identified (Control group, n = 66). All the parents had participated in a larger cohort study. The instruments used for measuring anxiety were STAI-state/trait, sense of coherence (SOC) and Parents' Expectations, Experiences, Reactions to an Ultrasound examination during pregnancy (PEER-U, State of Mind Index). Regarding the SOC and STAI-state/trait no significant differences were found between the cases and controls or within the respective group before and after the ultrasound examination. The cases had an increase in anxiety (more anxious) as measured by the instrument PEER-U after the examination, while the controls showed a significant better level of State of Mind Index (less anxious) after the examination, compared to before. Therefore PEER-U can be a more reliable instrument when studying state of mind (anxiety) in connection with ultrasound examinations, and as it is specific for this situation it does not appear to be time dependent.

Information for better or for worse: interviews with parents when their foetus was found to have choroid plexus cysts at a routine second trimester ultrasound.

The aim of the study was to gain a theoretical understanding of parents' experiences and handling of the situation, when their foetus was diagnosed as having choroid plexus cysts, at a routine second trimester ultrasound examination. Nine couples and one mother were interviewed using one open question. Analysis method was Grounded Theory. The main concern was anxiety and the core category became need for knowledge. The other categories were frightening and confusing, judging risk and making a choice and comforting. The parents felt information during the ultrasound examination was insufficient. The time delay between the diagnosis and the doctor's appointment was also often criticized. Most of the parents in this study wanted to know what can be diagnosed by ultrasound, even if there is a small risk that the child will have a malformation or chromosome abnormality. However, when the diagnosis is made, they need adequate information, otherwise unnecessary anxiety arises. By giving sufficient information without days of delay, anxiety can hopefully be minimized. Some written information was also requested. It is of utmost importance that the staff use the same terminology and the correct name of the soft marker to the parents.

Lipid metabolites as regulators of airway smooth muscle function.

Compelling evidence identifies airway smooth muscle (ASM) not only as a target but also a cellular source for a diverse range of mediators underlying the processes of airway narrowing and airway hyperresponsiveness in diseases such as asthma. These include the growing family of plasma membrane phospholipid-derived polyunsaturated fatty acids broadly characterised by the prostaglandins, leukotrienes, lipoxins, isoprostanes and lysophospholipids. In this review, we describe the enzymatic and non-enzymatic biosynthetic pathways of these lipid mediators and how these are influenced by drug treatment, oxidative stress and airways disease. Additionally, we outline their cognate receptors, many of which are expressed by ASM. We describe potential deleterious and protective roles for these lipid mediators in airway inflammatory and remodelling processes by describing their effects on diverse functions of ASM in asthma that have the potential to contribute to asthma pathogenesis and symptoms. These functions include contractile tone development, cytokine and extracellular matrix production, and cellular proliferation and migration.

Care during pregnancy and childbirth in Sweden: perspectives of lesbian women.

OBJECTIVE: to explore the views and experiences of care of lesbian women during pregnancy and childbirth. DESIGN, SETTING AND PARTICIPANTS: a qualitative study of 18 lesbian women in southern Sweden. FINDINGS: valid text units were formed through categorisation into four main categories: recognition of sexual orientation; openness; relationships within the homosexual family; and different encounters and attitudes within the health-care system. The interviewed women were positive about their care during pregnancy and childbirth. However, as in studies regarding women's experiences of care in general, lesbian women raised concerns about postnatal care, parent education and the structure of the patient records with no place for the female partner. KEY CONCLUSIONS AND IMPLICATIONS FOR PRACTICE: confirmation of parenthood was important, especially for the co-parent. The participants in this study felt that when they were open about their sexuality, this was met with an openness that they felt was confirming about their homsexuality. It is important for health-care providers not to make assumptions about women's sexuality.

Multivariate-activity mining for molecular quasi-species in a glutathione transferase mutant library.

A library of recombinant glutathione transferases (GSTs) generated by shuffling of DNA encoding human GST M1-1 and GST M2-2 was screened with eight alternative substrates, and the activities were subjected to multivariate analysis. Assays were made in lysates of bacteria in which the GST variants had been expressed. The primary data showed clustering of the activities in eight-dimensional substrate-activity space. For an incisive analysis, the rows of the data matrix, corresponding to the different enzyme variants, were individually scaled to unit length, thus accounting for different expression levels of the enzymes. The columns representing the activities with alternative substrates were subsequently individually normalized to unit variance and a zero mean. By this standardization, the data were adjusted to comparable orders of magnitude. Three molecular quasi-species were recognized by multivariate K-means and principal component analyses. Two of them encompassed the parental GST M1-1 and GST M2-2. A third one diverged functionally by displaying enhanced activities with some substrates and suppressed activities with signature substrates for GST M1-1 and GST M2-2. A fourth cluster contained mutants with impaired functions and was not regarded as a quasi-species. Sequence analysis of representatives of the mutant clusters demonstrated that the majority of the variants in the diverging novel quasi-species were structurally similar to the M1-like GSTs, but distinguished themselves from GST M1-1 by a Ser to Thr substitution in the active site. The data show that multivariate analysis of functional profiles can identify small structural changes influencing the evolution of enzymes with novel substrate-activity profiles.

Functionally diverging molecular quasi-species evolve by crossing two enzymes.

Molecular evolution is frequently portrayed by structural relationships, but delineation of separate functional species is more elusive. We have generated enzyme variants by stochastic recombinations of DNA encoding two homologous detoxication enzymes, human glutathione transferases M1-1 and M2-2, and explored their catalytic versatilities. Sampled mutants were screened for activities with eight alternative substrates, and the activity fingerprints were subjected to principal component analysis. This phenotype characterization clearly identified at least three distributions of substrate selectivity, where one was orthogonal to those of the parent-like distributions. This approach to evolutionary data mining serves to identify emerging molecular quasi-species and indicates potential trajectories available for further protein evolution.