Comparison of PD-L1 (22C3) Expression in Paired Primary and Metastatic Breast Carcinoma.

INTRODUCTION: PD-L1 immunohistochemistry (IHC) is being used as a predictive marker of the benefit derived from immunotherapy in several cancer types, including breast cancer. However, the insight gleaned of the prognostic and predictive value of PD-L1 status and its correlation with molecular characteristics during breast cancer progression remains limited. METHODS: We performed an PD-L1 (22C3) assay in pre-treatment primary and metastatic tumor sections from 33 patients with breast carcinoma, matched for post neoadjuvant chemotherapy (p-NACT). PD-L1 expression was evaluated using 3 scoring methods: immune cell (IC) and tumor cell (TC) with a 1% as the cutoff value, and combined positive scores (CPS) with a 1 as the cutoff value. Twenty-two samples from 11 patients had successful fluorescence in situ hybridization (FISH)-based molecular data available for analysis. RESULTS: In the 33 pre-treatment primary tumors, PD-L1 IC, TC, and CPS showed positive correlation with stromal tumor infiltrate lymphocytes (sTIL), histological grade 3, and triple negative breast carcinoma (TNBC). In the matched metastatic tumors, only PD-L1 IC showed a positive correlation with sTIL. The primary tumors showed a higher PD-L1 expression than the matched metastatic tumors by IC and CPS. Negative to positive conversion by CPS was identified in the metastatic tumors from lung, pleura and liver. p-NACT tumors also showed a trend of lower PD-L1 expression compared to the pre-treatment tumors. Six patients had matched samples for molecular and PD-L1 comparison, and none of them showed consistent gene alterations or PD-L1 expression among the primary, p-NACT and metastatic tumors. CONCLUSION: Our study showed a decrease in PD-L1 expression and disconnected molecular features during breast cancer progression. Repeating PD-L1 IHC testing could be considered in some specific metastatic sites if primary tumors were negative. Further studies are needed to identify other predictive factors for immune checkpoint inhibitor (ICI) therapy in patients with breast carcinoma.

Forest farming of shiitake mushrooms: an integrated evaluation of management practices.

Two outdoor shiitake (Lentinula edodes) cultivation experiments, established in Missouri USA in 1999 and 2000, produced mushrooms in 2000-2005. We examined shiitake production in response to substrate species, inoculum form, inoculum strain, and inoculation timing, using total mushroom weight per log as the primary response variable with log characteristics as covariates. The significantly greater mushroom weight produced by sugar maple logs compared with white or northern red oak was attributable to the higher proportion of undiscolored wood volume in the maple logs, rather than to bark thickness or log diameter. The "wide temperature range" shiitake strain produced significantly greater yield compared with the "warm" or "cold" weather strains. Both the wide-range and warm-weather strains were stimulated to fruit by significant rain events, while the cold-weather strain was responsive to temperature. Inoculation with sawdust spawn gave significantly greater yield than colonized wooden dowels or pre-packaged "thimble" plug inoculum. The second and third full years following inoculation were the most productive.

Exploration of the GM1 receptor-binding site of heat-labile enterotoxin and cholera toxin by phenyl-ring-containing galactose derivatives.

Cholera toxin (CT) and the closely related heat-labile enterotoxin of Escherichia coli (LT) are responsible for numerous cases of diarrhea worldwide, leading to considerable morbidity and mortality. The B subunits of these heterohexameric AB(5) toxins form a pentameric arrangement which is responsible for binding to the receptor GM1 of the target epithelial cells of the host. Blocking these B pentamer-receptor interactions forms an avenue for therapeutic intervention. Here, the structural characterization of potential receptor-blocking compounds are described based on the previously identified inhibitor m-nitrophenyl-alpha-D-galactoside (MNPG). The structure of a CTB-MNPG complex confirms that the binding mode of this inhibitor is identical in the two homologous toxins CT and LT and is characterized by a glycosyl linkage geometry that leads to displacement of a well ordered water molecule near the amide group of Gly33 by the O1-substituent of MNPG. This glycosyl geometry is not maintained in the absence of a substituent that can displace this water, as shown by a complex of LTB with p-aminophenyl-alpha-D-galactoside (PAPG). New compounds were synthesized to investigate the feasibility of maintaining the favorable binding interactions exhibited by MNPG while gaining increased affinity through the addition of hydrophobic substituents complementary to either of two hydrophobic regions of the receptor-binding site. The structural characterization of complexes of LTB with two of these compounds, 3-benzylaminocarbonylphenyl-alpha-D-galactoside (BAPG) and 2-phenethyl-7-(2,3-dihydrophthalazine-1,4-dione)-alpha-D-galactoside (PEPG), demonstrates a partial success in this goal. Both compounds exhibit a mixture of binding modes, some of which are presumably influenced by the local packing environment at multiple crystallographically independent binding sites. The terminal phenyl ring of BAPG associates either with the phenyl group of Tyr12 or with the hydrophobic patch formed by Lys34 and Ile58. The latter interaction is also made by the terminal phenyl substituent of PEPG, despite a larger ring system linking the galactose moiety to the terminal phenyl. However, neither BAPG nor PEPG displaces the intended target water molecule. Both of the designed compounds exhibit increased affinity relative to the galactose and to PAPG notwithstanding the failure to displace a bound water, confirming that additional favorable hydrophobic interactions can be gained by extending the starting inhibitor by a hydrophobic tail. The insight gained from these structures should allow the design of additional candidate inhibitors that retain both the glycosyl geometry and water displacement exhibited by MNPG and the favorable hydrophobic interactions exhibited by BAPG and PEPG.

Targeting adolescent mothers with depressive symptoms for early intervention.

Infants of mothers with depressive symptoms show developmental delays if symptoms persist over the first 6 months of the infant's life, thus highlighting the importance of identifying those mothers for early intervention. In Study 1, mothers with depressive symptoms (n = 160) and mothers without depressive symptoms (n = 100) and their infants were monitored to identify variables from the first 3 months that predict which mothers would still be symptomatic at 6 months. A "dysregulation" profile was noted for the infants of depressed mothers, including lower Brazelton scores, more indeterminate sleep, and elevated norepinephrine, epinephrine, and dopamine levels at the neonatal period, and greater right frontal EEG activation, lower vagal tone, and negative interactions at the 3- and 6-month periods. A group of maternal variables from the neonatal and 3-month assessments accounted for 51% of the variance in the mothers' continuing depressive symptoms. These variables included greater right frontal EEG activation, lower vagal tone, and less positive interactions at 3 months, and elevated norepinephrine, serotonin, and cortisol levels at the neonatal stage. In Study 2, a similar sample of mothers with depressive symptoms (n = 160) and without depressive symptoms (n = 100) was recruited and followed to 3 months. Those symptomatic mothers who had values above (or below) the median (depending on the negative direction) on the predictor variables identified in Study 1 (taken from the first 3 months) were then randomly assigned to an intervention or a control group at 3 months. These groups were then compared with each other, as well as with the group without depressive symptoms, at 6 and 12 months. The intervention, conducted from 3 to 6 months, consisted of free day care for the infants and a rehab program (social, educational, and vocational) plus several mood induction interventions for the mothers, including relaxation therapy, music mood induction, massage therapy, and mother-infant interaction coaching. Although the mothers who received the intervention continued to have more depressive symptoms than did the nondepressed mothers, their interactions significantly improved and their biochemical values and vagal tone normalized. Their infants also showed more positive interations, better growth, fewer pediatric complications, and normalized biochemical values, and by 12 months their mental and motor scores were better than those of the infants in the control group.

Structure of m-carboxyphenyl-alpha-D-galactopyranoside complexed to heat-labile enterotoxin at 1.3 A resolution: surprising variations in ligand-binding modes.

In the quest to develop drugs against traveller's diarrhoea and cholera, the structure of the B pentamer of heat-labile enterotoxin (LT) complexed with a new receptor-binding antagonist, m-carboxyphenyl-alpha-D-galactopyranoside, has been determined. The high resolution obtained for this structure allowed anisotropic refinement of the model. It was also now possible to confirm at a near-atomic resolution the structural similarity between the B subunits of LT and the closely related cholera toxin (CT), including the similarity in deviations of planarity of the same peptide unit in LT and CT. The structure of the LT complex clearly revealed different conformations for the m--carboxyphenyl moiety of the ligand in the five B subunits of LT, while the binding modes of the well defined galactopyranoside moieties were identical. In two binding sites the m-carboxyphenyl moiety displayed no significant electron density, demonstrating significant flexibility of this moiety. In a third binding site the m-carboxyphenyl moiety could be modelled unambiguously into the density. The two remaining binding sites were involved in crystal packing contacts and the density for the ligands in these two binding sites clearly revealed different binding modes, of which one conformation was identical to and one completely different from the conformation of m-carboxyphenyl-galactopyranoside in the third subunit. The multiple binding modes observed in the crystal may represent the ensemble of conformations of m-carboxyphenyl-alpha-D-galactopyranoside complexed to LT in solution.

Social networks for women with serious mental illness.

Assessment of ways to work with situations as they exist, and capitalizing on positive factors or strengths within the client's life context, may enhance the nurse-client relationship, and the likelihood of positive outcomes. It is important to identify "natural helpers" and "natural community resources," such as churches and adult centers within the community and to assess the actual and potential contributions and needs of these resources.

Living with serious mental illness: the desire for normalcy.

This article describes the desire for normalcy as experienced by 6 men and 13 women with serious mental illness based on secondary analysis of data from two previous qualitative studies. Original data were collected by structured and semi-structured interviews, and thematic analysis was done. Major categories of desire for normalcy are having normal things and experiences; doing meaningful activities; and being well, safe, free, and independent. Self-care actions to promote and maintain normalcy are identified, and finding are discussed in relation to Orem's self-care deficit theory of nursing. Implications for nursing theory, research, and practice are addressed.

Music shifts frontal EEG in depressed adolescents.

Recent studies have found that positive affect is associated with greater relative left frontal EEG activation and negative affect is associated with greater relative right frontal EEG activation. Further, chronically depressed adults typically display stable right frontal EEG activation. The present study investigated the effects of music on mood state and right frontal EEG activation associated with chronic depression. Fourteen chronically depressed female adolescents listened to rock music for a 23-minute session. These adolescents were compared with a control sample of chronically depressed female adolescents who were simply asked to sit and relax their minds and their muscles for the same time period. EEG was recorded during baseline, music, and postmusic for three minutes each, and saliva samples were collected before and after the session to determine the effects of the music on stress hormone (cortisol) levels. No group differences or changes were noted for observed or reported mood state. However, cortisol levels decreased and relative right frontal activation was significantly attenuated during and after the music procedure. It was concluded that music had positive effects on the physiological and biochemical measures even though observed and self-reported mood did not change.

Polydrug-using adolescent mothers and their infants receiving early intervention.

This study investigated the effects of an intervention for polydrug-using adolescent mothers. The program included educational, vocational, and parenting classes; social and drug rehab; and day care for their infants while they attended school half-day. The drug-exposed infants were similar to the nonexposed infants on traditional birth measures, although they had inferior Brazelton Neonatal Behavioral Assessment Scale scores, including habituation, orientation, abnormal reflexes, general irritability, and regulatory capacity. The drug-exposed infants also spent less time in quiet sleep and more time crying and showing stress behaviors. Both the mothers and the infants in the drug groups demonstrated inferior interactions, and their dopamine and serotonin levels were significantly higher. As early as 3 months (following 3 months of intervention), the drug rehab mothers and their infants looked more like the nondrug group in their interactions; by 6 months, they looked similar on virtually every measure. At 12 months, the infants of drug rehab mothers (versus the drug control group) had superior Early Social Communication Scale scores and Bayley Mental scale scores, as well as significantly greater head circumference and fewer pediatric complications. The drug rehab mothers also improved on several lifestyle variables. They demonstrated a lower incidence of continued drug use and repeat pregnancy, and a greater number continued school, received a high school or general equivalency diploma, or were placed in a job. Thus, a relatively cost-effective high school based intervention had positive effects on both adolescent mothers who had used drugs and their infants.

PIP: The impact of an early childhood intervention program on polydrug-abusing US adolescent mothers and their infants was evaluated. The program, which was located in a vocational school attended by the mothers, included drug rehabilitation, social skills training, parenting classes, job training, and relaxation therapy. Outcomes in 126 drug-exposed mothers 16-21 years of age who participated in the program were compared to those recorded among non-drug-using adolescent mothers who participated in the program and drug-using control mothers who did not participate. All three groups were similar in terms of age, education, socioeconomic status, and ethnicity, but drug-abusing mothers had higher rates of depression and stress. At baseline, drug-exposed infants had lower scores on the measures of habituation, orientation, abnormal reflexes, general irritability, and regulatory capacity on the Neonatal Behavioral Assessment Scale. Drug-exposed infants spent less time sleeping and more time crying and showing stress behaviors. The drug groups also had lower Optimal Interaction Rating Scale scores for both mothers and infants. Their dopamine and serotonin levels were higher than those recorded among non-drug-using mothers and their cortisol levels were lower. However, after 6 months of participation in the intervention program, the drug-using mothers had Beck Depression Inventory scores and interaction ratings that approached those of non-drug-using mothers and exceeded those among drug-using controls. Similar trends were observed for infants' head circumference and scores on the Early Social Communication Scale and the Bayley Mental Status Scale. Moreover, drug-using adolescent mothers who participated in the program demonstrated a lower incidence of repeat pregnancy and continued drug use than those who were not enrolled in the program; moreover, they were more likely to receive their high school diploma and be placed in jobs. Interventions such as this have the potential to attenuate the developmental delays of infants of drug-exposed adolescents.

Formal and informal care of people with psychiatric disorders: historical perspectives and current trends.

People with severe and persistent mental illness have a broad array of needs related to health and well being. In the present era of shrinking health care resources and managed care, it is not likely that the formal health care system can or will be sufficiently responsive to the needs of this population. To address adequately the care and treatment of individuals with serious mental illness, it is essential to identify and use all available resources, both in the formal health care system and within informal systems of care.

The effect of retrieval cues on visual preferences and memory in infancy: evidence for a four-phase attention function.

Bahrick and Pickens (1995) proposed a four-phase model of infant attention, suggesting that recent memories are expressed as a visual preference for novelty, intermediate memories as a null preference, and remote memories as a preference for familiarity. The present study tested a hypothesis generated from this model that a retrieval cue would increase memory accessibility and shift visual preferences toward greater novelty to resemble more recent memories. Results confirmed our predictions. After retention intervals associated with remote memory, previously observed familiarity preferences shifted to null preferences, whereas after a retention interval associated with intermediate memory, the previously observed null preference shifted to a novelty preference. Further, a second experiment found that increasing the exposure to the retrieval cue could shift the familiarity preference to a novelty preference. These findings support the four-phase model of infant attention and suggest that novelty, null, and familiarity preferences lie along a continuum and shift as a function of memory accessibility.

EEG stability in infants/children of depressed mothers.

The stability of EEG was examined in infants of depressed (BDI > 16) and non-depressed mothers (BDI < 9) from 3 months to 3 years. Of the 32 infants seen at 3 months, 15 were seen again at 3 years of age. Seven of the eight children who had exhibited right frontal EEG asymmetry as infants still showed that EEG asymmetry pattern at the 3 year visit. Children with right frontal EEG asymmetry at 3 years were observed to be more inhibited during an exploratory play task, and children of depressed versus non-depressed mothers were less empathetic during simulated maternal distress.

Massage therapy reduces anxiety and enhances EEG pattern of alertness and math computations.

Twenty-six adults were given a chair massage and 24 control group adults were asked to relax in the massage chair for 15 minutes, two times per week for five weeks. On the first and last days of the study they were monitored for EEG, before, during and after the sessions. In addition, before and after the sessions they performed math computations, they completed POMS Depression and State Anxiety Scales and they provided a saliva sample for cortisol. At the beginning of the sessions they completed Life Events, Job Stress and Chronic POMS Depression Scales. Group by repeated measures and post hoc analyses revealed the following: 1) frontal delta power increased for both groups, suggesting relaxation; 2) the massage group showed decreased frontal alpha and beta power (suggesting enhanced alertness); while the control group showed increased alpha and beta power; 3) the massage group showed increased speed and accuracy on math computations while the control group did not change; 4) anxiety levels were lower following the massage but not the control sessions, although mood state was less depressed following both the massage and control sessions; 5) salivary cortisol levels were lower following the massage but not the control sessions but only on the first day; and 6) at the end of the 5 week period depression scores were lower for both groups but job stress score were lower only for the massage group.

Depressed mothers' touching increases infants' positive affect and attention in still-face interactions.

The effects of depressed mothers' touching on their infants' behavior were investigated during the still-face situation. 48 depressed and nondepressed mothers and their 3-month-old infants were randomly assigned to control and experimental conditions. 4 successive 90-sec periods were implemented: (A) normal play, (B) still-face-no-touch, (C) still-face-with-touch, and (A) normal play. Depressed and nondepressed mothers were instructed and shown how to provide touch for their infants during the still-face-with-touch period. Different affective and attentive responses of the infants of depressed versus the infants of nondepressed mothers were observed. Infants of depressed mothers showed more positive affect (smiles and vocalizations) and gazed more at their mothers' hands during the still-face-with-touch period than the infants of nondepressed mothers, who grimaced, cried, and gazed away from their mothers' faces more often. The results suggest that by providing touch stimulation for their infants, the depressed mothers can increase infant positive affect and attention and, in this way, compensate for negative effects often resulting from their typical lack of affectivity (flat facial and vocal expressions) during interactions.

Preschoolers of dysphoric mothers.

Dysphoric mothers rated their preschool children as having more internalizing and externalizing behavior problems than children of nondysphoric mothers. Observers rated the dysphoric mothers as having more negative affect during play interactions, although their children's affect was not rated more negatively. The dysphoric mother-infant dyads were also rated as having a poorer quality interaction. These data suggest that mothers' chronic dysphoria (75% were chronically dysphoric) has a negative impact on the mothers' perceptions of their children as well as the mothers' and children's interaction behavior.

Infant memory for object motion across a period of three months: implications for a four-phase attention function.

Memory for object motion in 3-month-old infants was investigated across retention intervals of 1 or 3 months in three studies using a novelty preference method. Following familiarization to an object undergoing one of two types of motion, visual preferences for the novel motion were assessed after retention intervals of 1 min, 1 day, and 1 month (Experiment 1, N = 120) and 1 min, 1 day, 2 weeks, and 1 month (Experiment 2, N = 74). Results of both studies indicated a significant preference for the novel motion at the 1-min delay, a significant preference for the familiar motion at the 1-month delay, and no preferences at the intermediate retention intervals. In Experiment 3, memory was assessed after a 3-month interval and again, a significant familiarity preference was obtained. These results demonstrate that memory for object motion lasts across retention intervals of 1 and 3 months and that novelty and familiarity preferences interact with retention time. A four-phase function relating visual preferences and retention time was proposed. Phase 1, recent memory, is characterized by a novelty preference; phase 2, intermediate memory, is a period of transition characterized by no visual preference; phase 3, remote memory, is characterized by a familiarity preference; and phase 4, inaccessible memory, is also characterized by no preference. The finding of a transition period at intermediate retention times suggests that null preferences should not necessarily be taken as evidence of forgetting. Rather, more extended retention intervals should be included to interpret null findings obtained in the novelty preference method.

In vitro model of human umbilical venous perfusion to study the effects of meconium staining of the umbilical cord.

Because meconium diffuses into placental and umbilical cord tissue, its bioactive components are potentially capable of entering the fetal circulation. Meconium diluted in Krebs solution produces vasoconstriction in human umbilical vein segments that are suspended in a milieu similar to that of the amniotic cavity. Based on published observations, we wanted to determine if very diluted meconium could induce vasospasm in intact umbilical cords. Human umbilical cords were obtained from full-term deliveries, cut into 2 equal segments, cannulated, and perfused continuously as test and control cords. The study included four parts: (1) intraluminal meconium (n = 6; 0.01-1% solution); (2) 4-hour exposure to 10% meconium-stained amniotic fluid of intact umbilical cords (n = 8); (3) 4-hour exposure to a 10% meconium-stained amniotic fluid of umbilical cords denuded of their epithelial layer (n = 5); (4) 24-hour exposure to a 10% meconium-stained amniotic fluid of intact umbilical cords (n = 9). Significant vasoconstriction of the umbilical vein occurred only with the intraluminal meconium solution exposure (p = 0.034). In this in vitro model meconium staining of the umbilical cord was not associated with vasoconstriction of the umbilical vein.

The effects of extended perioperative pentoxifylline on random skin flap survival.

PURPOSE: This study was designed to determine the effects of long-term perioperative pentoxifylline administration on random skin flap survival in an appropriate animal model. A secondary objective was to document bioavailability of pentoxifylline in the animal model by measuring blood levels of parent compound and metabolites at regular intervals and comparing these to levels measured in humans. MATERIALS AND METHODS: A randomized control study of the effects of oral pentoxifylline on the survival of "random" skin flaps was conducted at the animal care facility of an academic tertiary referral center on six randomly selected Yorkshire pigs. Oral pentoxifylline was administered daily to four pigs for 3 months, and two pigs received placebo. Pentoxifylline blood levels for each experimental animal were measured after 4, 8, and 12 weeks of daily dosing. Blood viscosity, fibrinogen, and hematocrit were measured for each of the six animals on day 1, day 30, day 60, and day 91. On day 91, 12 surgical random skin flaps were elevated on each of the six animals and immediately sewn back in the donor bed. Pentoxifylline dosing was continued for 2 weeks, and placebo was continued in control animals. On postoperative day 15, all animals were killed and all flaps were measured individually for area of viability. The outcome measure was the detection of statistically significant increase in survival area in skin flaps of those animals administered perioperative pentoxifylline. RESULTS: No significant augmentation of flap survival was noted in experimental animals when compared with those in the control group; no significant difference was noted between or within groups of experimental animals. Pentoxifylline and metabolite blood levels in all experimental animals at every interval were noted to be comparable to those documented in human studies; metabolite concentrations conformed to expected patterns as observed in humans. No significant alterations of blood viscosity, fibrinogen, or hematocrit were measured in the experimental animals when compared with those in the control group. The only animal showing mean flap survival outside the 95% confidence interval was one in the control group. CONCLUSION: No augmentation of random skin flap survival could be shown in the pig model after a 3 month regimen of daily oral pentoxifylline. Blood levels of pentoxifylline in experimental animals were compared with those documented in humans. No alteration of blood viscosity, fibrinogen, or hematocrit was noted in any of the experimental animals when compared with each other or with those in the control group.

"Depressed" mothers' perceptions of their preschool children's vulnerability.

"Depressed" mothers who scored in the depression range on the Beck Depression Inventory when their infants were 3-months-old were asked to complete the Vulnerable Child Scale when their children were preschool age. Depressed versus non-depressed mothers rated their preschool children as being more vulnerable (as did an independent observer) as well as having more behavior problems. The vulnerability ratings were significantly related to infancy stage measures including the POMS (mothers' depressed mood), the infants' behavior ratings and heart rate range during interactions with a nondepressed stranger. These measures of the mothers' and infants' behavior tapped during early infancy may have contributed to the mothers having perceived their children as "vulnerable" during the preschool stage.

Current concepts of the lethal midline granuloma syndrome.

"Lethal midline granuloma" is an unusual but devastating spectrum of lymphoproliferative disorders that is now thought to include four entities: idiopathic midline destructive disease, Wegener's granulomatosis, polymorphic reticulosis, and nasal lymphoma. These disorders must be considered in the differential diagnosis of any patient with chronic, nonspecific complaints localized to the upper airway. An aggressive diagnostic and therapeutic approach may significantly decrease morbidity and death resulting from these lesions.